Christopher Parish

Selective localisation of neuro-specific T lymphocytes in the central nervous system

Publisher: Elsevier BV

Date: 04-1992

DOI: 10.1016/0165-5728(92)90008-9

Abstract: Using experimental autoimmune encephalomyelitis (EAE) in the rat as a model of central nervous system (CNS) inflammation, activated and quiescent T lymphocytes with different antigen specificities were labelled with the fluorescent dye Hoechst 33342 and tested by fluorescence microscopy for their ability to accumulate in different regions of the spinal cord and in other organs at varying times post inoculation. With this highly sensitive assay it was found that activated myelin basic protein (MBP)-specific T cell lines accumulated in the spinal cord (a 1000-fold increase in the lumbar/sacral region by day 4) and caused clinical signs of EAE. In contrast, interleukin-2 (IL-2)-maintained (quiescent) MBP-specific T cell lines failed to accumulate in the CNS and cause disease. Activated ovalbumin (OA)-specific and purified protein derivative of tuberculin (PPD)-specific T cell lines were also found at significantly higher levels in the spinal cord than non-activated cells although they failed to accumulate to a substantial degree when injected alone. When injected with activated MBP-specific T cells the activated OA- and PPD-specific cell lines accumulated in the spinal cord following initial accumulation of the MBP-specific cells, demonstrating that during the inflammatory process there is considerable non-specific recruitment of cells into the inflammatory site. CNS accumulation of activated MBP-specific T cell lines occurred 1-2 days later in irradiated animals than in non-irradiated recipients. This was consistent with irradiated animals also exhibiting a later onset of disease and suggests that irradiation may directly affect the endothelium in a way that makes it less adhesive. In conclusion, this study demonstrates that activated lymphocytes of any specificity enter the spinal cord, and that the neuro-antigen specific cells accumulate there and lead to the recruitment of other cells. Non-activated cells, even those with neural antigen specificity fail to enter the cord. Understanding the nature of what an 'activated' lymphocyte is may allow us to design strategies to inhibit such immune-mediated inflammation.

Activation of cathepsin D by glycosaminoglycans

Publisher: Wiley

Date: 03-04-2023

DOI: 10.1111/J.1742-4658.2009.07444.X

Abstract: We have previously shown that heparin can increase the activity of the proenzyme form of Alzheimer's beta-site amyloid precursor protein cleaving enzyme 1 (BACE1). Cathepsin D (CD) is a member of the aspartic protease family and has sequence similarity to BACE1. Therefore, we examined whether heparin and other glycosaminoglycans (GAGs) can influence the activity of CD. Heparin and other GAGs were found to stimulate the activity of recombinant proCD. Desulfation of heparin almost abolished the stimulation, indicating that sulfate groups were important for the stimulatory effect. In addition, the stimulation was dependent on the length of the GAG chain, as larger GAGs were more potent in their ability to stimulate proCD than shorter fragments. In the presence of heparin, limited autocatalytic proteolysis of the proenzyme was increased, suggesting that heparin increases the activity of proCD by accelerating the conversion of proCD, which has little activity, to pseudoCD, an active form lacking residues 1-26 of the prodomain. Furthermore, the activity of spleen-derived mature CD, which lacks the entire 44 amino acid residue prodomain, was also increased by heparin, indicating that the catalytic domain of CD contains at least one region to which GAGs bind and stimulate enzyme activity. Because heparin also stimulated the activity of pseudoCD, proenzyme activation was probably accelerated by the interaction of heparin with the catalytic domain of pseudoCD. However, it is possible that heparin may also activate the proenzyme directly. On the basis of this study, we propose that GAGs may regulate CD activity in vivo.

Immunology and cell biology publication of the Year Award 2009

Publisher: Wiley

Date: 10-2010

DOI: 10.1038/ICB.2010.102

Bystander B cells rapidly acquire antigen receptors from activated B cells by membrane transfer (Proceedings of the National Academy of Sciences of the United States of America (2008) 105, 11, (4259-4264) DOI: 10.1073/pnas. 0800259105))

Publisher: Proceedings of the National Academy of Sciences

Date: 25-08-2009

DOI: 10.1073/PNAS.0908246106

Detection and isolation of an Ia glycoprotein antigen from mouse serum

Publisher: Springer Science and Business Media LLC

Date: 12-1977

DOI: 10.1007/BF01575666

Histidine-rich glycoprotein binds heparanase and regulates its enzymatic activity and cell surface interactions

Publisher: Elsevier BV

Date: 09-2010

DOI: 10.1016/J.BIOCEL.2010.05.008

Abstract: Heparanase, an endo-beta-D-glucuronidase, is involved in numerous normal physiological and pathological processes, such as inflammation, wound healing and tumour metastasis/angiogenesis, through its ability to mediate the degradation of heparan sulfate, a key structural component of the extracellular matrix and on the surface of cells. Identifying endogenous molecules that can regulate heparanase activity will aid the understanding of its molecular function in health and disease and provide the potential for development of novel anti-cancer and anti-inflammatory therapeutics. The ability of the extracellular heparanase to tether onto cell surface heparan sulfate proteoglycans and other receptor(s), such as the cation-independent mannose-6-phosphate receptor, is key to its activation, function and uptake into intracellular compartments. Here we describe experiments demonstrating that a relatively abundant plasma glycoprotein, histidine-rich glycoprotein, directly interacts with platelet-derived heparanase and enhances its enzymatic activity. The findings in this study also show that histidine-rich glycoprotein interferes with heparanase binding to cell surface receptors, particularly heparan sulfate proteoglycans. Thus, the interaction between histidine-rich glycoprotein and heparanase can potentially regulate the role of heparanase in a variety of physiological and pathological conditions.

Cloning of the prometastatic enzyme heparanase

Publisher: Elsevier BV

Date: 04-2000

DOI: 10.1016/S0753-3322(00)89055-1

A basement‐membrane permeability assay which correlates with the metastatic potential of tumour cells

Publisher: Wiley

Date: 30-09-1992

DOI: 10.1002/IJC.2910520309

Abstract: We describe an in vitro assay for measuring the ability of tumour cells to permeabilize basement membranes, using transwell chambers coated with the reconstituted basement membrane, matrigel. Unlike previous matrigel-based procedures which quantified passage of tumour cells across a matrigel barrier, the new assay measures the ability of tumour cells to degrade the basement membrane and increase the diffusion rate of fluorescent (FL) dextran through the barrier. The procedure has the major advantage that permeability can be rapidly and accurately quantified, either by fluorometry or by the use of radiolabelled dextran, thus avoiding tedious and subjective scoring methods. Optimal conditions for the assay are described. In addition, it is demonstrated that the assay can clearly discriminate between metastatic and non-metastatic tumour cell lines, metastatic tumours permeabilizing the basement membrane and non-metastatic counterparts failing to do so. A range of enzyme inhibitors suggested that the increase in basement-membrane permeability caused by the metastatic mammary adenocarcinoma 13762 MAT is probably dependent upon the synergistic action of several degradative enzymes, namely proteases, type-IV collagenase, and heparanase. Furthermore, the ability to permeabilize the basement membrane was dependent upon intact tumour cells tumour cell extracts, lysates and supernatants were inactive.

Exploring science pedagogy on the web 2.0/mobile border: Teachers’ views of a mobile wiki-based inquiry approach

Publisher: Modestum Ltd

Date: 09-01-2023

DOI: 10.29333/EJMSTE/12820

Abstract: This paper reports on teachers’ experiences of a 10-week, wiki-based, predominantly mobile science education intervention for secondary school students. It was designed to shift traditional pedagogical approaches towards more inquiry-based approaches by combining the strengths of wikis and mobile smart devices. The four participating Macau science teachers, with shared responsibility for 250 students, noted greater student engagement and reported some shifts towards more constructivist pedagogy and more active student learning. Evidence emerged of the value of personalization, collaboration, online feedback, and authentic learning, but the lack of mobile optimization of the wiki platform negatively impacted the degree of personalization, collaboration and authenticity. Lessons were learned about the need for educational technology interventions to carefully consider the intersection between software and hardware the reasons for the recent decline of wikis as educational tools and the ongoing educational value of web 2.0-style collaboration in our increasingly mobile era.

A novel fluorescent-based assay reveals that thrombopoietin signaling and Bcl-XL influence, respectively, platelet and erythrocyte lifespans

Publisher: Elsevier BV

Date: 06-2010

DOI: 10.1016/J.EXPHEM.2010.03.003

Abstract: The factors determining platelet and erythrocyte lifespan are not completely understood, despite extensive study. The lack of success may be attributed to the methods used to measure lifespan kinetics, all of which require processing of cells prior to analysis, and the inconsistent and potentially inappropriate use of mathematical models for data analysis. The aims of this study were to establish an in vivo platelet and erythrocyte labeling method using carboxyfluorescein diacetate succinimidyl ester (CFSE), determine the most appropriate mathematical model for lifespan analysis, and apply both to the study of factors that control platelet and erythrocyte lifespans. Control, c-mpl knockout (KO), and Bcl-X(L) mutant mice were injected with CFSE and platelet and erythrocyte fluorescence followed over time. Datasets were analyzed using linear, exponential, multiple-hit, and lognormal mathematical models. In vivo CFSE labeling of platelets and erythrocytes requires no postcollection processing, proved stable, nontoxic, nonimmunogenic, and the lifespans were highly reproducible. Mathematical modeling revealed the lognormal model gave a robust fit to control and extreme datasets when either extrinsic or intrinsic factors determined lifespan. Using these methods, platelet lifespans were found to be significantly shortened in thrombopoietin-receptor-deficient mice independent of blood loss, and the antiapoptotic protein Bcl-X(L) was shown to play a role in prolonging erythrocyte lifespans. The simultaneous study of platelet and erythrocyte lifespans using in vivo CFSE labeling with lognormal modeling yielded insight into common intrinsic and extrinsic platelet and erythrocyte lifespan determinants and provides an improved methodology for use in this field of research.

Use of an in vivo FTA assay to assess the magnitude, functional avidity and epitope variant cross-reactivity of T cell responses following HIV-1 recombinant poxvirus vaccination

Publisher: Public Library of Science (PLoS)

Date: 29-08-2014

DOI: 10.1371/JOURNAL.PONE.0105366

Intracellular fluorescent labelling of cells for analysis of lymphocyte migration

Publisher: Elsevier BV

Date: 11-1984

DOI: 10.1016/0022-1759(84)90364-8

Abstract: A procedure for analysing in vivo migration of lymphocytes labelled in vitro using intracellular fluorochromes is described. Comparison of carboxyfluorescein diacetate, a cytoplasmic label, with Hoechst dye No. 33342 (H33342), a DNA-binding fluorochrome, indicated that H33342 is superior. The concentration of H33342 used for labelling does not significantly affect viability or lymphocyte migration and permits long-term visualization. H33342 allows quantitation of in vivo migration in cell suspension and histological localization in frozen sections. Fluorescence is retained in fixed frozen sections for at least 3 months. This method can be used for analyses of lymphocyte migration and maturation.

Structural organization and polypeptide composition of the avian adenovirus core

Publisher: American Society for Microbiology

Date: 11-1984

DOI: 10.1128/JVI.52.2.638-649.1984

Abstract: CELO virus (fowl adenovirus 1) contained three core polypeptides of molecular weights 20,000, 12,000, and 9,500. The core was similar to that of human adenoviruses, with some evidence of compact subcore domains. Micrococcal nuclease digestion of CELO virus cores produced a smear of DNA fragments of gradually decreasing size, with no nucleosome subunit or repeat pattern. Moreover, when digested cores were analyzed without protease treatment, there was again no evidence of a nucleosome substructure neither DNA fragments nor core proteins entered a 4% polyacrylamide gel. The organization of the core is thus quite unlike that of chromatin. Restriction endonuclease analysis of the DNA from digested cores showed that the right end was on the outside of the core. We suggest that adenovirus DNA is condensed into the core by cross-linking and neutralization by the core proteins, beginning with the packaging sequence at the center of the core and ending with the right end of the DNA on the outside.

Use of sulfated linked cyclitols as heparan sulfate mimetics to probe the heparin/heparan sulfate binding specificity of proteins

Publisher: Elsevier BV

Date: 03-2005

DOI: 10.1074/JBC.M410769200

Carbohydrate-Defined Antigens Controlled by the I Region

Publisher: Springer US

Date: 1981

DOI: 10.1007/978-1-4684-3758-4_11

A Novel Method for Monitoring Tropical Cyclones’ Movement Using GNSS Zenith Tropospheric Delay

Publisher: MDPI AG

Date: 23-06-2023

DOI: 10.3390/RS15133247

Abstract: Precipitable water vapor (PWV) is an important meteorological factor for predicting extreme weather events such as tropical cyclones, which can be obtained from zenith tropospheric delay (ZTD) by using a conversion. A time difference of ZTD arrival (TDOZA) model was proposed to monitor the movement of tropical cyclones, and the fifth-generation reanalysis dataset of the European Centre for Medium-range Weather Forecasting (ERA5)-derived ZTD (ERA5-ZTD) was used to estimate the movement of tropical cyclones based on the model. The global navigation satellite system-derived ZTD and radiosonde data-derived PWV (RS-PWV) were used to test the accuracy of the ERA5-ZTD and analyze the correlation between ZTD and PWV, respectively. The statistics showed that the mean Bias, RMS and STD of the ERA5-ZTD were 6.4 mm, 17.1 mm and 16.5 mm, respectively, and the mean correlation coefficient of the ERA5-ZTD and RS-PWV was 0.951, which indicates that the ZTD can be used to predict weather events instead of PWV. Then, spatiao-temporal characteristics of ZTD during the four tropical cyclone (i.e., Merbok, ROKE, Neast and Hato) periods in 2017 were analyzed, and the result showed that the moving directions of ZTD and the tropical cyclones were consistent. Thus, the ZTD time series over the ERA5 grids around the tropical cyclones’ paths were used to estimate the velocity of the tropical cyclones based on the TDOZA model, when the tropical cyclones are approaching or leaving. Compared with the result from the China Meteorological Administration, the mean absolute and relative deviations of the TDOZA model-derived velocity were 2.55 km/h and 10.0%, respectively. These results suggest that ZTD can be used as a new supplementary meteorological parameter for monitoring tropical cyclone events.

Histidine-rich glycoprotein: The Swiss Army knife of mammalian plasma

Publisher: American Society of Hematology

Date: 17-02-2011

DOI: 10.1182/BLOOD-2010-09-303842

Abstract: Histidine-rich glycoprotein (HRG), also known as histidine-proline-rich glyco-protein, is an abundant and well-characterized protein of vertebrate plasma. HRG has a multidomain structure that allows the molecule to interact with many ligands, including heparin, phospholipids, plasminogen, fibrinogen, immunoglobulin G, C1q, heme, and Zn2+. The ability of HRG to interact with various ligands simultaneously has suggested that HRG can function as an adaptor molecule and regulate numerous important biologic processes, such as immune complex/necrotic cell athogen clearance, cell adhesion, angiogenesis, coagulation, and fibrinolysis. The present review covers the proposed multifunctional roles of HRG with a focus on recent findings that have led to its emergence as a key regulator of immunity and vascular biology. Also included is a discussion of the striking functional similarities between HRG and other important multifunctional proteins found in plasma, such as C-reactive protein, C1q, β2 glycoprotein I, and thrombospondin-1.

A Rapid and Accurate Bioluminescence-Based Migration Assay Permitting Analysis of Tumor Cell/Stromal Cell Interactions

Publisher: MDPI AG

Date: 23-01-2020

DOI: 10.3390/MPS3010010

Abstract: Bioluminescent tumor cell lines are used extensively in vivo to monitor tumor growth and metastasis but rarely used in vitro to follow tumor cell behavior. Tumor cell migration is frequently studied in vitro using transwell assays, however, current methods do not permit the co-incubation of tumor cells with different stromal cell types for analysis of the effects of intercellular cross-talk on tumor cell migration. We describe a novel migration assay using bioluminescent tumor cell lines that is rapid, accurate, and permits the study of the effects of tumor cell-stromal cell interactions on tumor cell migratory behavior.

Serological analysis of antigen-specific helper factors specific for poly-L(Tyr, Glu)-poly-DLAla--poly-LLys [(T, G)-A--L] and L Glu60-LAla30-LTyr10 (GAT).

Publisher: Wiley

Date: 07-1979

DOI: 10.1002/EJI.1830090703

Abstract: In vitro prepared antigen-specific helper factors reactive to the synthetic polypeptide antigens poly-L(Tyr, Glu)-poly-DLAla--poly-LLys [(T, G)-A--L] or LGlu60-LAla30-LTyr10 (GAT) and bearing Ia determinants were analyzed serologically to determine the nature of the Ia determinants they expressed. I subregion-specific mouse anti-Ia antisera were used, and showed that (T, G)-A--L-specific helper factor (HF) contains I-A subregion-controlled determinants, whereas GAT-specific HF carries I-J subregion-controlled antigens. This unexptected finding was confirmed in both the H-2k and H-2 b haplotypes, using a variety of anti-I-J antisera. Rabbit anti-Ia antisera also reacted with both HF which raised the possibility that the Ia determinants on HF may be carbohydrate in nature. The fact that HF has a low molecular weight and yet contains Ia determinants, antigen-binding capacity and idiotypic markers is compatible with this interpretation.

Cancer immunotherapy: The past, the present and the future

Publisher: Wiley

Date: 04-2003

DOI: 10.1046/J.0818-9641.2003.01151.X

Abstract: One of the most controversial issues in immunology for over a century has been whether an effective immune response can be elicited against malignant tumours. Whether the immunology community has believed cancer immunotherapy is feasible or impossible has been largely determined by the prevailing immunological paradigms at that time. In fact, during the last 110 years it is possible to trace at least five dramatic fluctuations in attitude towards cancer immunotherapy. It now appears, however, that overwhelming evidence is available to support the view that both the innate and adaptive immune responses can recognize and eliminate tumours. On the other hand, it remains to be seen if these immune responses can be harnessed to control cancer as, at the time of diagnosis, many tumours have already been immunoselected to be highly resistant to immune elimination. Based on these observations it is argued that immunotherapy approaches, other than the generation of tumour-specific cytotoxic T lymphocytes, must be explored. Alternative strategies include recruiting tumouricidal myeloid cells into tumours, generating antiangiogenic immune responses and directing innate immunity to hypoxia-induced ligands on tumour cells.

Low zone tolerance to bacterial flagellin in adult rats: a possible role for antigen localized in lymphoid follicles.

Publisher: Proceedings of the National Academy of Sciences

Date: 10-1968

DOI: 10.1073/PNAS.61.2.556

Abstract: Child-staff ratios are a key quality indicator in early childhood education and care (ECEC) programs. Better ratios are believed to improve child outcomes by increasing opportunities for in idual interactions and educational instruction from staff. The purpose of this systematic review, and where possible, meta-analysis, was to evaluate the association between child-staff ratios in preschool ECEC programs and children's outcomes. Searches of Medline, PsycINFO, ERIC, websites of large datasets and reference sections of all retrieved articles were conducted up to July 3, 2015. Cross-sectional or longitudinal studies that evaluated the relationship between child-staff ratios in ECEC classrooms serving preschool aged children and child outcomes were independently identified by two reviewers. Data were independently extracted from included studies by two raters and differences between raters were resolved by consensus. Searches revealed 29 eligible studies (31 s les). Child-staff ratios ranged from 5 to 14.5 preschool-aged children per adult with a mean of 8.65. All 29 studies were included in the systematic review. However, the only meta-analysis that could be conducted was based on three studies that explored associations between ratios and children's receptive language. Results of this meta-analysis were not significant. Results of the qualitative systematic review revealed few significant relationships between child-staff ratios and child outcomes construed broadly. Thus, the available literature reveal few, if any, relationships between child-staff ratios in preschool ECEC programs and children's developmental outcomes. Substantial heterogeneity in the assessment of ratios, outcomes measured, and statistics used to capture associations limited quantitative synthesis. Other methodological limitations of the research integrated in this synthesis are discussed.

Fractionation of detergent lysates of cells by ammonium sulphate-induced phase separation

Publisher: Elsevier BV

Date: 08-1986

DOI: 10.1016/0003-2697(86)90284-8

Abstract: A procedure is described for fractionating detergent lysates of cells based on the ability of (NH4)2SO4 to induce phase separation of detergents such as Triton X-100, sodium deoxycholate, and sodium cholate, into detergent-rich and detergent-depleted phases. An analysis of six murine lymphocyte cell surface molecules revealed that the partitioning in Triton X-100 of each molecule was highly dependent upon the (NH4)2SO4 concentration, each antigen partitioning into the detergent-rich phase at a defined salt concentration. In contrast, none of the six molecules appeared in the detergent-rich phase of a Triton X-114 phase separation, even though two of the molecules, namely Ly-2/3 and L3T4, are well-characterized integral membrane proteins. It was also observed that (NH4)2SO4 resulted in the partitioning of many nonmembrane proteins into the detergent-rich phase, indicating that the procedure can be used to fractionate all cellular proteins. By judicious choice of (NH4)2SO4 concentrations, precipitation of cellular proteins at two different (NH4)2SO4 concentrations, and combining the method with subcellular fractionation prior to detergent solubilization, substantial enrichment and concentration of particular cellular proteins could be achieved.

Interactions with Asialo-Glycoprotein Receptors and Platelets Are Dispensable for CD8+ T Cell Localization in the Murine Liver

Publisher: The American Association of Immunologists

Date: 15-06-2022

DOI: 10.4049/JIMMUNOL.2101037

Abstract: Liver-resident CD8+ T cells can play critical roles in the control of pathogens, including Plasmodium and hepatitis B virus. Paradoxically, it has also been proposed that the liver may act as the main place for the elimination of CD8+ T cells at the resolution of immune responses. We hypothesized that different adhesion processes may drive residence versus elimination of T cells in the liver. Specifically, we investigated whether the expression of asialo-glycoproteins (ASGPs) drives the localization and elimination of effector CD8+ T cells in the liver, while interactions with platelets facilitate liver residence and protective function. Using murine CD8+ T cells activated in vitro, or in vivo by immunization with Plasmodium berghei sporozoites, we found that, unexpectedly, inhibition of ASGP receptors did not inhibit the accumulation of effector cells in the liver, but instead prevented these cells from accumulating in the spleen. In addition, enforced expression of ASGP on effector CD8+ T cells using St3GalI-deficient cells lead to their loss from the spleen. We also found, using different mouse models of thrombocytopenia, that severe reduction in platelet concentration in circulation did not strongly influence the residence and protective function of CD8+ T cells in the liver. These data suggest that platelets play a marginal role in CD8+ T cell function in the liver. Furthermore, ASGP-expressing effector CD8+ T cells accumulate in the spleen, not the liver, prior to their destruction.

The use of carboxyfluorescein diacetate succinimidyl ester (CFSE) to monitor lymphocyte proliferation

Publisher: MyJove Corporation

Date: 12-10-2010

DOI: 10.3791/2259

Nuclear import of early growth response-1 involves importin-7 and the novel nuclear localization signal serine-proline-serine

Publisher: Elsevier BV

Date: 06-2011

DOI: 10.1016/J.BIOCEL.2011.03.004

Abstract: A three amino acid sequence, Ser/Thr-Pro-Ser/Thr, was recently identified and characterized as a novel nuclear localization signal (Chuderland et al., 2008). The immediate-early gene product, early growth response-1 is a three zinc finger containing transcription factor implicated in a wide variety of pathologies, and has a bipartite nuclear localization domain identified two decades ago. Efficient nuclear localization of Egr-1 is vital to its function as a transcription factor. Interestingly, Egr-1 also contains a C-terminal SPS domain (residues 482-484 in murine Egr-1). We hypothesized that (482)SPS(484) may also serve as a novel nuclear localization signal in Egr-1. We found that this sequence directs Egr-1 to the nucleus in transfected Chinese hamster ovary cells and show by co-immunoprecipitation analysis that Egr-1 forms a complex with importin-7. (482)SPS(484) is required for Egr-1's interaction with importin-7. Moreover, importin-7 knockdown with RNAi showed that Egr-1 nuclear translocation is importin-7-dependent. This study demonstrates that the nuclear translocation of Egr-1 is partially dependent on (482)SPS(484) and involves importin-7, and sheds light on the molecular mechanisms regulating the cellular localization of this pathophysiologically important transcription factor.

The immunomodulatory compound 2-acetyl-4-tetrahydroxybutyl imidazole causes sequestration of lymphocytes in non-lymphoid tissues

Publisher: Wiley

Date: 10-1997

DOI: 10.1038/ICB.1997.77

Abstract: 2-Acetyl-4(5)-(1,2,3,4-tetrahydroxybutyl) imidazole (THI) is an immunomodulatory compound which causes a reversible lymphopenia in mice by an unknown mechanism. In this study, we investigated the whereabouts of cells lost from the blood and the spleen during THI treatment Homing studies following is injection of fluorescently labelled splenocytes into THI-pretreated recipients showed that THI increased labelled cells in the liver, lungs and kidneys of THI-treated mice. Furthermore, the sequestration in the liver occurred just 1.5 h after injection of labelled cells with the increase still being present at 24 h after injection. Microscopic examination of liver sections indicated that fluorescent lymphocytes were clustered within the liver sinusoids in THI-treated mice, possibly associated with endothelial cells. The liver retention of lymphocytes was confirmed by immunohistochemical studies which showed a significant increase of T cells in the liver of THI-treated mice. To determine the subset of lymphocytes which are lost from the spleen and sequestered in non-lymphoid organs, lymphocytes remaining in the spleen after THI treatment were characterized. Our results confirmed that THI reduced B cells, CD4+ and CD8+ T cells and cells expressing CD62L, CD44 and IL-2R in the spleen.

BCLA CLEAR - Evidence-based contact lens practice

Publisher: Elsevier BV

Date: 04-2021

DOI: 10.1016/J.CLAE.2021.02.008

Mitogens and T-independent antigens stimulate T lymphocytes to secrete Ia antigens

Publisher: Elsevier BV

Date: 09-1977

DOI: 10.1016/0008-8749(77)90141-1

Abstract: The effect of antibiotic exposure of phenotypically smooth gram-negative bacteria on binding by the human lipid A-reactive monoclonal antibody HA-1A (trademark of Centocor, Inc.) was examined by liquid-phase immunoassay and by dual-parameter flow cytometry (fluorescence-activated cell sorter [FACS]) analysis. HA-1A exhibited dose-dependent binding to untreated rough gram-negative bacteria such as the Escherichia coli D21F2 Re chemotype strain but little binding to untreated smooth strains such as E. coli O111:B4, or to gram-positive bacteria. However, overnight incubation of E. coli O111:B4 with inhibitory concentrations of ceftazidime produced dose-dependent enhancement of HA-1A binding. Similar augmentation of HA-1A binding was observed when other smooth strains were exposed to cell wall-active agents. Dual-parameter FACS analysis of E. coli O111:B4 exposed overnight to two times the MIC of ceftazidime revealed a decrease in forward light scatter, indicating a reduction in average cell size or bacterial fragmentation, accompanied by a striking increase in lipid A-inhibitable HA-1A binding. Moreover, ceftriaxone, but not gentamicin, produced a marked increase in propidium iodide uptake, indicating an increase in bacterial cell permeability, and a corresponding enhancement of HA-1A binding. Antibiotic-induced enhancement of HA-1A binding to smooth strains of gram-negative bacteria thus appears related to specific alterations in bacterial cell morphology resulting in exposure of the epitope recognized by HA-1A.

A Novel Mechanism for Complement Activation at the Surface of B Cells Following Antigen Binding

Publisher: The American Association of Immunologists

Date: 15-10-2006

DOI: 10.4049/JIMMUNOL.177.8.5155

Abstract: Coligation of CD21 with BCR on the surface of B cells provides a costimulatory signal essential for efficient Ab responses to T-dependent Ags. To achieve this, Ag must be directly linked to C3 fragments, but how this occurs in vivo is not fully understood. Using BCR transgenic mice, we demonstrated that C3 was deposited on the surface of B cells following both high- and moderate-affinity Ag binding. This was dependent on the specific binding of IgM to the BCR-bound Ag and can occur independently of soluble immune complex formation. Based on these data, we propose a novel model in which immune complexes can form directly on the surface of the B cell following Ag binding. This model has implications for our understanding of B lymphocyte activation.

Fluorescent dyes for lymphocyte migration and proliferation studies

Publisher: Wiley

Date: 12-1999

DOI: 10.1046/J.1440-1711.1999.00877.X

Abstract: Fluorescent dyes are increasingly being exploited to track lymphocyte migration and proliferation. The present paper reviews the properties and performance of some 14 different fluorescent dyes that have been used during the last 20 years to monitor lymphocyte migration. Of the 14 dyes discussed, two stand out as being the most versatile in terms of long-term tracking of lymphocytes and their ability to quantify lymphocyte proliferation. They are the intracellular covalent coupling dye carboxyfluorescein diacetate succinimidyl ester (CFSE) and the membrane inserting dye PKH26. Both dyes have the advantage that they can be used to track cell ision, both in vitro and in vivo, due to the progressive halving of the fluorescence intensity of the dyes in cells after each ision. However, CFSE appears to have the edge over PKH26 based on homogeneity of lymphocyte staining and cost. Two other fluorescent dyes, although not suitable for lymphocyte proliferation studies, are valuable tracking dyes for short-term (up to 3 day) lymphocyte migration experiments, namely the DNA-binding dye Hoechst 33342 and the cytoplasmic dye calcein. In the future it is highly likely that additional fluorescent dyes, with different spectral properties to CFSE, will become available, as well as membrane inserting fluorescent dyes that more homogeneously label lymphocytes than PKH26.

Histidine-rich glycoprotein: A novel adaptor protein in plasma that modulates the immune, vascular and coagulation systems

Publisher: Wiley

Date: 04-2005

DOI: 10.1111/J.1440-1711.2005.01320.X

Abstract: Histidine-rich glycoprotein (HRG) is an abundant plasma glycoprotein that has a multidomain structure, interacts with many ligands, and has been shown to regulate a number of important biological processes. HRG ligands include Zn(2+) and haem, tropomyosin, heparin and heparan sulphate, plasminogen, plasmin, fibrinogen, thrombospondin, IgG, FcgammaR and complement. In many cases, the histidine-rich region of the molecule enhances ligand binding following interaction with Zn(2+) or exposure to low pH, conditions associated with sites of tissue injury or tumour growth. The multidomain nature of HRG indicates that it can act as an extracellular adaptor protein, bringing together disparate ligands, particularly on cell surfaces. HRG binds to most cells primarily via heparan sulphate proteoglycans, binding which is also potentiated by elevated free Zn(2+) levels and low pH. Recent reports have shown that HRG can modulate angiogenesis and additional studies have shown that it may regulate other physiological processes such as cell adhesion and migration, fibrinolysis and coagulation, complement activation, immune complex clearance and phagocytosis of apoptotic cells. This review outlines the molecular, structural, biological and clinical properties of HRG as well as describing the role of HRG in various physiological processes.

The Contribution of Neutrophils and NETs to the Development of Type 1 Diabetes

Publisher: Frontiers Media SA

Date: 06-07-2022

DOI: 10.3389/FIMMU.2022.930553

Abstract: Type 1 diabetes (T1D) is an autoimmune disease resulting from the destruction of insulin-producing beta cells in pancreatic islets. T lymphocytes are the claimed pathogenic effectors but abnormalities of other immune cell types, including neutrophils, also characterize T1D development. During human T1D natural history, neutrophils are reduced in the circulation, while accumulate in the pancreas where release of neutrophil extracellular traps (NETs), or NETosis, is manifest. Recent-onset T1D patients also demonstrate activated circulating neutrophils, associated with a unique neutrophil gene signature. Neutrophils can bind to platelets, leading to the formation of platelet-neutrophil aggregates (PNAs). PNAs increase in the circulation during the development of human T1D and provide a mechanism for neutrophil activation and mobilization/recruitment to the pancreas. In non-obese diabetic or NOD mice, T1D autoimmunity is accompanied by dynamic changes in neutrophil numbers, activation state, PNAs and/or NETosis/NET proteins in the circulation, pancreas and/or islets. Such properties differ between stages of T1D disease and underpin potentially indirect and direct impacts of the innate immune system in T1D pathogenesis. Supporting the potential for a pathogenic role in T1D, NETs and extracellular histones can directly damage isolated islets in vitro , a toxicity that can be prevented by small polyanions. In human T1D, NET-related damage can target the whole pancreas, including both the endocrine and exocrine components, and contribute to beta cell destruction, providing evidence for a neutrophil-associated T1D endotype. Future intervention in T1D could therefore benefit from combined strategies targeting T cells and accessory destructive elements of activated neutrophils.

Demonstration of lymphocyte surface lectins that recognize sulphated polysaccharides

Publisher: The Company of Biologists

Date: 04-1984

DOI: 10.1242/JCS.67.1.145

Abstract: Cholate extracts of murine lymphocytes were shown to contain haemagglutinating activity against autologous erythrocytes. The species specificity and sugar inhibition pattern of the haemagglutinin closely paralleled the specificity of autorosetting, an interaction that had been shown previously to involve the recognition by lymphocytes of carbohydrate structures on autologous erythrocytes. The probable identity of the haemagglutinin and autorosetting receptors was confirmed by experiments utilizing the unique plasma protein autorosette inhibition factor, which appears to block both interactions by masking carbohydrate acceptor sites on erythrocytes. Detailed sugar inhibition studies revealed that the haemagglutinin and autorosetting receptors have a high affinity for certain sulphated polysaccharides, such as heparin and dextran sulphate. Since similar sulphated polysaccharides have been shown previously to inhibit lymphocyte recirculation, a possible role for these receptors in lymphocyte homing and recirculation is discussed.

A simple and rapid acrylamide gel method for estimating the molecular weights of proteins and protein subunits

Publisher: Elsevier BV

Date: 04-1970

DOI: 10.1016/0003-2697(70)90128-4

Abstract: High levels of HIV risk behaviors and prevalence have been reported among Puerto Rican people who inject drugs (PRPWID) since early in the HIV epidemic. Advances in HIV prevention and treatment have reduced HIV among people who inject drugs (PWID) in the United States. We examined HIV-related data for PRPWID in Puerto Rico and the US Northeast to assess whether disparities continue. Injection drug use as a risk for HIV is still overrepresented among Puerto Ricans. Lower availability of syringe exchanges, drug abuse treatment, and antiretroviral treatment for PWID in Puerto Rico contribute to higher HIV risk and incidence. These disparities should be addressed by the development of a federally supported Northeast-Puerto Rico collaboration to facilitate and coordinate efforts throughout both regions.

Human platelet heparanase: Purification, characterization and catalytic activity

Publisher: Portland Press Ltd.

Date: 15-03-1998

DOI: 10.1042/BJ3301341

Abstract: Heparan sulphate (HS) is an important component of the extracellular matrix (ECM) and the vasculature basal lamina (BL) which functions as a barrier to the extravasation of metastatic and inflammatory cells. Platelet-tumour cell aggregation at the capillary endothelium results in activation and degranulation of platelets. Cleavage of HS by endoglycosidase or heparanase activity produced in relatively large amounts by the platelets and the invading cells may assist in the disassembly of the ECM and BL, and thereby facilitate cell migration. Using a recently published rapid, quantitative assay for heparanase activity towards HS [Freeman, C. and Parish, C. R. (1997), Biochem. J., 325, 229-237], human platelet heparanase has now been purified 1700-fold to homogeneity in 19% yield by a five column procedure, which consists of concanavalin A-Sepharose, Zn2+-chelating-Sepharose, Blue A-agarose, octyl-agarose and gel filtration chromatography. The enzyme, which was shown to be an endoglucuronidase that degrades both heparin and HS, has a native molecular mass of 50 kDa when analysed by gel filtration chromatography and by SDS/PAGE. Platelet heparanase degraded porcine mucosal HS in a stepwise fashion from a number average molecular mass of 18.5 to 13, to 8 and finally to 4.5 kDa fragments as determined by gel filtration analysis. Bovine lung heparin was degraded from 8.9 to 4.8 kDa while porcine mucosal heparin was degraded from 8.1 kDa to 3.8 and finally to 2.9 kDa fragments. Studies of the enzyme's substrate specificity using modified heparin analogues showed that substrate cleavage required the presence of carboxyl groups, but O- and N-sulphation were not essential. Inhibition studies demonstrated an absolute requirement for the presence of O-sulphate groups. Platelet heparanase was inhibited by heparin analogues which also inhibited tumour heparanase, suggesting that sulphated polysaccharides which inhibit tumour metastasis may act to prevent both tumour cell and platelet heparanase degradation of endothelial cell surface HS and the basal laminar.

Synthesis and biological evaluation of some enantiomerically pure C8cC15 monoseco analogues of the phenanthroquinolizidine-type alkaloids cryptopleurine and julandine

Publisher: CSIRO Publishing

Date: 2008

DOI: 10.1071/CH08190

Abstract: A series of enantiomerically pure C8c–C15 monoseco analogues, 23–30, of the alkaloids cryptopleurine (1) and julandine (2) have been prepared using cinnamyl chloride 37 and (S)- or (R)-2-methylpiperidine as key building blocks. Two related compounds, 31 and 32, have also been synthesized. Each of these analogues has been subjected to various biological evaluations and most of them show dramatically reduced cytotoxicity compared with parent system 1. Nevertheless, they are potent anti-angiogenic agents. The formation and single-crystal X-ray analysis of the spirocyclic dienone 54, a by-product arising from attempts to prepare analogue 32, is also described.

Heparan sulfate and inflammation

Publisher: Springer Science and Business Media LLC

Date: 09-2005

DOI: 10.1038/NI0905-861

Inhibition of Plasmodium falciparum growth in vitro and adhesion to chondroitin-4-sulfate by the heparan sulfate mimetic PI-88 and other sulfated oligosaccharides

Publisher: American Society for Microbiology

Date: 08-2006

DOI: 10.1128/AAC.00313-06

Abstract: A panel of sulfated oligosaccharides was tested for antimalarial activity and inhibition of adhesion to the placental malaria receptor chondroitin-4-sulfate (CSA). The heparan sulfate mimetic PI-88, currently undergoing phase II anticancer trials, displayed the greatest in vitro antimalarial activity against Plasmodium falciparum (50% inhibitory concentration of 7.4 μM) and demonstrated modest adhesion inhibition to cell surface CSA.

Lack of a correlation between cell-mediated immunity to the carrier and the carrier-hapten helper effect

Publisher: Rockefeller University Press

Date: 03-1974

DOI: 10.1084/JEM.139.3.779

Abstract: The relationship between cell-mediated immunity to the carrier and the carrier-hapten helper effect was studied in the rat by using three forms of the carrier which differed in their capacity to induce carrier-specific delayed-type hypersensitivity. The three carriers were polymerized flagellin (POL), flagellin (FIN), and acetoacetylated flagellin (AFIN), which induced FIN-specific delayed-type hypersensitivity in the order AFIN & FIN & POL. Helper cells for the anti-DNP antibody responses to a range of DNP-FIN conjugates appeared to be almost inversely related to cell-mediated immunity to the carrier, being in the order POL & FIN ⪖ AFIN. These differences occurred whether the carriers were injected in saline or FCA, but were less pronounced with the heavily DNP-conjugated flagellins.

The protective capacities of fractionated immune thoracic duct lymphocytes against Nippostrongylus brasiliensis.

Publisher: Elsevier BV

Date: 04-1978

DOI: 10.1016/0008-8749(78)90172-7

Fibrin exposure triggers αIIbβ3‐independent platelet aggregate formation, ADAM10 activity and glycoprotein VI shedding in a charge‐dependent manner

Publisher: Elsevier BV

Date: 06-2020

DOI: 10.1111/JTH.14797

Histidine-rich glycoprotein binds to human IgG and Clq and inhibits the formation of insoluble immune complexes

Publisher: American Chemical Society (ACS)

Date: 06-1997

DOI: 10.1021/BI962573N

A functional heparan sulfate mimetic implicates both heparanase and heparan sulfate in tumor angiogenesis and invasion in a mouse model of multistage cancer

Publisher: Springer Science and Business Media LLC

Date: 04-04-2005

DOI: 10.1038/SJ.ONC.1208602

Abstract: Heparan sulfate proteoglycans are integral components of the extracellular matrix that surrounds all mammalian cells. In addition to providing structural integrity, they act as a storage depot for a variety of heparan sulfate (HS)-binding proteins, including growth factors and chemokines. Heparanase is a matrix-degrading enzyme that cleaves heparan sulfate side chains from the core proteoglycans, thus liberating such HS-binding proteins, as well as potentially contributing to extracellular matrix degradation. Here, we report that heparanase mRNA and protein expression are increased in the neoplastic stages progressively unfolding in a mouse model of multistage pancreatic islet carcinogenesis. Notably, heparanase is delivered to the neoplastic lesions in large part by infiltrating Gr1+/Mac1+ innate immune cells. A sulfated oligosaccharide mimetic of heparan sulfate, PI-88, was used to inhibit simultaneously both heparanase activity and HS effector functions. PI-88 had significant effects at distinct stages of tumorigenesis, producing a reduction in the number of early progenitor lesions and an impairment of tumor growth at later stages. These responses were associated with decreased cell proliferation, increased apoptosis, impaired angiogenesis, and a substantive reduction in the number of invasive carcinomas. In addition, we show that the reduction in tumor angiogenesis is correlated with a reduced association of VEGF-A with its receptor VEGF-R2 on the tumor endothelium, implicating heparanase in the mobilization of matrix-associated VEGF. These data encourage clinical applications of inhibitors such as PI-88 for the many human cancers where heparanase expression is elevated or mobilization of HS-binding regulatory factors is implicated.

Histidine-rich glycoprotein binding to T-cell lines and its effect on T-cell substratum adhesion is strongly potentiated by zinc

Publisher: Wiley

Date: 06-1996

DOI: 10.1111/J.1365-2567.1996.TB00005.X

Abstract: Histidine-rich glycoprotein (HRG), a plasma protein that binds heparin and alent cations, has been implicated in immune regulation through its ability to modulate complement function, macrophage Fc receptor expression and phagocytosis, and its ability to inhibit the proliferation of human peripheral blood T cells in vitro. In the present work we used fluorescence flow cytometry to study the binding of human HRG to the human T-cell lines Jurkat and MT4, and to the murine antigen-specific T-cell clone D10, and to study the effect of alent cations zinc and copper on this binding. Our results show that HRG binds strongly to these cell lines at 4 degrees, and that the binding is markedly potentiated by physiological concentrations of zinc (20 microM), and to a lesser extent by copper (10 microM). In contrast to previous studies, HRG binding was largely inhibited by 50 micrograms/ml heparin, both in the absence and in the presence of zinc, suggesting that HRG interacts primarily through glycosaminoglycans on the T-cell surface. Studies using confocal fluorescence microscopy indicated that following incubation of MT4 cells with HRG in the presence of zinc at 4 degrees, the HRG was localized exclusively at the plasma membrane, but was actively internalized after incubation at 37 degrees. Interestingly, HRG interfered with the ability of D10 cells to adhere to tissue culture plastic, as well as to laminin-, collagen- or fibronectin-coated culture dishes. This effect was markedly potentiated by 20 microM zinc, and was partially reversed by heparin. The results suggest that zinc markedly potentiates the binding of HRG to T cells, and that HRG and zinc may play an important role in regulating the adhesion of T cells to other cells and the extracellular matrix.

Low-molecular-weight Ia antigens in normal mouse serum

Publisher: Springer Science and Business Media LLC

Date: 12-1976

DOI: 10.1007/BF01576974

Histidine-rich glycoprotein regulates the binding of monomeric IgG and immune complexes to monocytes

Publisher: Oxford University Press (OUP)

Date: 08-1999

DOI: 10.1093/INTIMM/11.8.1275

Abstract: Histidine-rich glycoprotein (HRG) is a relatively abundant plasma protein which we have shown previously inhibits the formation of insoluble immune complexes (IC). In this study we examined the ability of HRG to regulate the binding of monomeric IgG and IC to monocytes. Initial studies demonstrated that HRG interacts with FcgammaRI on the monocytic cell line THP1 and blocks the binding of monomeric IgG to these cells. However, despite totally blocking the binding of monomeric IgG to FcgammaRI, pre-incubation of THP1 cells with HRG had no effect on the binding of IC to these cells. In contrast, depending on the HRG:IgG molar ratio, pre-incubation of monomeric IgG with HRG resulted in either enhanced or reduced IgG binding to FcgammaRI. Similarly, under certain highly defined conditions, incorporation of HRG in IgG-containing IC potentiated the binding of IC to THP1 cells. The key conditions involved incorporating approximately equimolar concentrations of HRG and IgG in the IC, the IC being formed at a near equivalence antigen:antibody ratio and usually physiological concentration (20 microM) of Zn(2+) being present. Collectively these observations indicate that HRG is an important regulator of IC uptake by monocytes. Thus HRG can interact with FcgammaRI on monocytes and block monomeric IgG binding, whereas when incorporated in IgG containing IC, HRG can enhance the uptake of IC by monocytes, probably via its heparan sulfate binding domain.

A rapid quantitative assay for the detection of mammalian heparanase activity

Publisher: Portland Press Ltd.

Date: 07-1997

DOI: 10.1042/BJ3250229

Abstract: Heparan sulphate (HS) is an important component of the extracellular matrix and the vasculature basal laminar which functions as a barrier to the extravasation of metastatic and inflammatory cells. Cleavage of HS by endoglycosidase or heparanase activity produced by invading cells may assist in the disassembly of the extracellular matrix and basal laminar, and thereby facilitate cell migration. Heparanase activity has previously been shown to be related to the metastatic potential of murine and human melanoma cell lines [Nakajima, Irimura and Nicolson (1988) J. Cell. Biochem. 36, 157–167]. To determine heparanase activity, porcine mucosal HS was partially de-N-acetylated and re-N-acetylated with [3H]acetic anhydride to yield a radiolabelled substrate. This procedure prevented the masking of, or possible formation of, new heparanase-sensitive cleavage sites as has been observed with previous methods of radiolabelling. Heparanase activity in a variety of tissues and cell homogenates including human platelets, colonic carcinoma cells, umbilical vein endothelial cells and rat mammary adenocarcinoma cells (both metastatic and non-metastatic variants) and liver homogenates all degraded the substrate in a stepwise fashion from 18.5 to approximately 13, 8 and finally to 4.5 kDa fragments, as assessed by gel-filtration analysis, confirming the substrate as suitable for the detection of heparanase activity present in a variety of cells and tissues. A rapid quantitative assay was developed with the HS substrate using a novel method for separating degradation products from the substrate by taking advantage of the decreased affinity of the heparanase-cleaved products for the HS-binding plasma protein chicken histidine-rich glycoprotein (cHRG). Incubation mixtures were applied to cHRG–Sepharose columns, with unbound material corresponding to heparanase-degradation products. Heparanase activity was determined for a variety of human, rat and murine cell and tissue homogenates. The highly metastatic rat mammary adenocarcinoma and murine lung carcinoma cell lines had four to ten times the heparanase activity of non-metastatic variants, confirming the correlation of heparanase activity with metastatic potential. Human cancer patients had twice the serum heparanase levels of normal healthy adults. The assay will be valuable for the determination of heparanase activity from a variety of tissue and cell sources, as a diagnostic tool for the determination of heparanase potential, and for the development of specific inhibitors of heparanase activity and metastasis.

Are murine marginal‐zone macrophages the splenic white pulp analog of high endothelial venules?

Publisher: Wiley

Date: 11-1995

DOI: 10.1002/EJI.1830251127

Abstract: The entry of lymphocytes into the spleen, in contrast to lymph nodes, does not involve high endothelial venule (HEV) interaction. The precise point of entry, as well as the mechanism by which lymphocytes enter the lymphoid areas of the spleen, remains controversial. We examined in detail the effect of two agents, pertussis toxin (PT) and the sulfated polysaccharide fucoidan, on splenic lymphocyte entry and positioning. These have previously been shown to interfere with lymphocyte extravasation across HEV. PT prevents lymphocyte extravasation, but not binding, to HEV, whereas fucoidan prevents binding and thus subsequent extravasation. Studies presented here show that pretreatment of murine lymphocytes with PT does not numerically affect entry into spleen, but profoundly alters lymphocyte positioning within the spleen. When fluorescently labeled, PT-treated lymphocytes are injected intravenously, they initially accumulate in the marginal zone, in apparent association with the layer of marginal zone macrophages (MZM phi) which form a shell around the white pulp. They fail to traverse this layer into the white pulp, and subsequently localize in the red pulp. In contrast, untreated cells initially appear in the marginal zone, then continue to migrate into the white pulp after traversing the MZM phi layer. The localization of PT-pretreated lymphocytes adjacent to the MZM phi layer is disrupted by intravenous administration of fucoidan. Using a flow cytometric assay of aggregation between MZM phi and lymphocytes, we confirmed that fucoidan is also able to inhibit this association in vitro, whereas PT has no effect on this interaction. We propose that MZM phi in the mouse are the splenic analog of HEV, forming the port of entry of lymphocytes into the white pulp of the spleen.

Drug-induced thrombocytopenia: Development of a novel NOD/SCID mouse model to evaluate clearance of circulating platelets by drug-dependent antibodies and the efficacy of IVIG

Publisher: American Society of Hematology

Date: 16-09-2010

DOI: 10.1182/BLOOD-2010-02-268326

Abstract: Drug-induced immune thrombocytopenia (DITP) is an adverse drug effect mediated by drug-dependent antibodies. Intravenous immunoglobulin (IVIG) is frequently used to treat DITP and primary immune thrombocytopenia (ITP). Despite IVIG's proven beneficial effects in ITP, its efficacy in DITP is unclear. We have established a nonobese diabetic/severe combined immunodeficient (NOD/SCID) mouse model of DITP in which human platelets survive for more than 24 hours, allowing platelet clearance by DITP/ITP antibodies to be studied. Rapid human platelet clearance was uniformly observed with all quinine-induced thrombocytopenia (QITP) patient sera studied (mean platelet lifespans: QITP 1.5 ± 0.3 hours vs controls 16.5 ± 4.3 hours), consistent with the clinical presentation of DITP. In contrast, clearance rates with ITP antibodies were more variable. IVIG treatment partially prevented platelet clearance by DITP and ITP antibodies. Our results suggest that the NOD/SCID mouse model is useful for investigating the efficacy of current and future DITP therapies, an area in which there is little experimental evidence to guide treatment.

A sensitive rosetting method for detecting subpopulations of lymphocytes which react with alloantisera.

Publisher: Elsevier BV

Date: 04-1978

DOI: 10.1016/0022-1759(78)90254-5

Abstract: A procedure is described for directly estimating the proportion of mouse lymphoid cell suspensions which react with alloantisera. The method entails reacting Ig-capped lymphoid cells with alloantisera, and then assessing the uptake of alloantibodies by rosetting the lymphocytes with SRBC coated with sheep IgG specific for mouse Ig. This rosetting procedure was found to be generally more sensitive than the conventional dye exclusion microcytotoxicity test for detecting the binding of alloantibodies to lymphocytes. Furthermore, the rosette method has the advantage that, unlike the complement lysis technique, it has a low and reproducible background and lymphocyte subpopulations which react with alloantisera can be isolated.

Synthesis and preliminary evaluation of 5,7-dimethyl-2-aryl-3H-pyrrolizin-3-ones as angiogenesis inhibitors.

Publisher: Elsevier BV

Date: 04-2016

DOI: 10.1016/J.BMCL.2016.02.033

Abstract: Sunitinib (Sutent®) is a receptor tyrosine kinase (RTK) and angiogenesis inhibitor approved for the treatment of renal cell carcinomas, gastrointestinal stromal tumours and pancreatic neuroendocrine tumours. A key structural motif retained throughout medicinal chemistry efforts during sunitinib's development was the indoline-2-one group. In the search for new anti-angiogenic scaffolds, we previously reported that non-indoline-2-one-based derivatives of semaxanib (SU5416, a structurally simpler sunitinib predecessor that underwent Phase III trials) are active as angiogenesis inhibitors, indicating that the group is not essential for activity. This Letter describes the synthesis and structure-activity relationships of another class of non-indoline-2-one angiogenesis inhibitors related to sunitinib/semaxanib the 5,7-dimethyl-2-aryl-3H-pyrrolizin-3-ones. A focussed library of 19 analogues was prepared using a simple novel process, wherein commercially available substituted arylacetic acids activated with an amide coupling reagent (HBTU) were reacted with the potassium salt of 3,5-dimethyl-1H-pyrrole-2-carbaldehyde in one-pot. Screening of the library using a cell-based endothelial tube formation assay identified 6 compounds with anti-angiogenesis activity. Two of the compounds were advanced to the more physiologically relevant rat aortic ring assay, where they showed similar inhibitory effects to semaxanib at 10μg/mL, confirming that 5,7-dimethyl-2-aryl-3H-pyrrolizin-3-ones represent a new class of angiogenesis inhibitors.

Protein determination on an automatic spectrophotometer

Publisher: Elsevier BV

Date: 03-1982

DOI: 10.1016/0003-2697(82)90578-4

Immunology and cell biology's impact factor rises for the fourth consecutive year

Publisher: Wiley

Date: 08-2010

DOI: 10.1038/ICB.2010.97

Regulation of the immune response by antibody. I. Suppression of antibody formation and concomitant enhancement of cell-mediated immunity by passive antibody

Publisher: Elsevier BV

Date: 05-1972

DOI: 10.1016/0008-8749(72)90006-8

Abstract: Given the predominance of invasive fungal disease (IFD) amongst the non-immunocompromised adult critically ill population, the potential benefit of antifungal prophylaxis and the lack of generalisable tools to identify high risk patients, the aim of the current study was to describe the epidemiology of IFD in UK critical care units, and to develop and validate a clinical risk prediction tool to identify non-neutropenic, critically ill adult patients at high risk of IFD who would benefit from antifungal prophylaxis. Data on risk factors for, and outcomes from, IFD were collected for consecutive admissions to adult, general critical care units in the UK participating in the Fungal Infection Risk Evaluation (FIRE) Study. Three risk prediction models were developed to model the risk of subsequent Candida IFD based on information available at three time points: admission to the critical care unit, at the end of 24 h and at the end of calendar day 3 of the critical care unit stay. The final model at each time point was evaluated in the three external validation s les. Between July 2009 and April 2011, 60,778 admissions from 96 critical care units were recruited. In total, 359 admissions (0.6 %) were admitted with, or developed, Candida IFD (66 % Candida albicans). At the rate of candidaemia of 3.3 per 1000 admissions, blood was the most common Candida IFD infection site. Of the initial 46 potential variables, the final admission model and the 24-h model both contained seven variables while the end of calendar day 3 model contained five variables. The end of calendar day 3 model performed the best with a c index of 0.709 in the full validation s le. Incidence of Candida IFD in UK critical care units in this study was consistent with reports from other European epidemiological studies, but lower than that suggested by previous hospital-wide surveillance in the UK during the 1990s. Risk modeling using classical statistical methods produced relatively simple risk models, and associated clinical decision rules, that provided acceptable discrimination for identifying patients at 'high risk' of Candida IFD. The FIRE Study was reviewed and approved by the Bolton NHS Research Ethics Committee (reference: 08/H1009/85), the Scotland A Research Ethics Committee (reference: 09/MRE00/76) and the National Information Governance Board (approval number: PIAG 2-10(f)/2005).

Demonstration of carbohydrate- and protein determined Ia antigens by monoclonal antibodies.

Publisher: Springer Science and Business Media LLC

Date: 12-1980

DOI: 10.1007/BF01567815

Abstract: Management of tuberculosis (TB) has witnessed several changes over the past decades. While medical management is now the mainstay of therapy, surgical intervention was once the only treatment option physicians had to offer. We discuss some historical surgical procedures and take a quick glance at the evolution of TB therapy. We note the importance of adequate history-taking and the implications of what seemingly obsolete techniques may have in contemporary practice. We also highlight the re-emergence of surgical options in the modern era with the rise of multidrug-resistance.

Bacterial Flagellin as an Antigen and Immunogen

Publisher: Springer US

Date: 1972

DOI: 10.1007/978-1-4757-1343-5_3

Computational analyses of the catalytic and heparin-binding sites and their interactions with glycosaminoglycans in glycoside hydrolase family 79 endo-d-glucuronidase (heparanase)

Publisher: Oxford University Press (OUP)

Date: 11-07-2012

DOI: 10.1093/GLYCOB/CWR095

Abstract: Mammalian heparanase is an endo-β-glucuronidase associated with cell invasion in cancer metastasis, angiogenesis and inflammation. Heparanase cleaves heparan sulfate proteoglycans in the extracellular matrix and basement membrane, releasing heparin/heparan sulfate oligosaccharides of appreciable size. This in turn causes the release of growth factors, which accelerate tumor growth and metastasis. Heparanase has two glycosaminoglycan-binding domains however, no three-dimensional structure information is available for human heparanase that can provide insights into how the two domains interact to degrade heparin fragments. We have constructed a new homology model of heparanase that takes into account the most recent structural and bioinformatics data available. Heparin analogs and glycosaminoglycan mimetics were computationally docked into the active site with energetically stable ring conformations and their interaction energies were compared. The resulting docked structures were used to propose a model for substrates and conformer selectivity based on the dimensions of the active site. The docking of substrates and inhibitors indicates the existence of a large binding site extending at least two saccharide units beyond the cleavage site (toward the nonreducing end) and at least three saccharides toward the reducing end (toward heparin-binding site 2). The docking of substrates suggests that heparanase recognizes the N-sulfated and O-sulfated glucosamines at subsite +1 and glucuronic acid at the cleavage site, whereas in the absence of 6-O-sulfation in glucosamine, glucuronic acid is docked at subsite +2. These findings will help us to focus on the rational design of heparanase-inhibiting molecules for anticancer drug development by targeting the two heparin/heparan sulfate recognition domains.

Editorial: Carbohydrates: The yet to be tasted sweet spot of immunity

Publisher: Frontiers Media SA

Date: 17-06-2015

DOI: 10.3389/FIMMU.2015.00314

Dependence of the adaptive immune response on innate immunity: Some questions answered but new paradoxes emerge

Publisher: Wiley

Date: 12-1997

DOI: 10.1038/ICB.1997.83

Abstract: Recently a new model of vertebrate immunity has been gaining popularity. In this new model it is hypothesized that activation of innate immunity is a prerequisite for an adaptive immune response to an antigen. Following activation the innate system induces key costimulator molecules on APC, which are essential for antigen-driven clonal expansion of T and B cells. The model largely explains the need for adjuvants in the induction of adaptive immunity, provides a possible mechanism for the immune system to perceive the biological nature of a pathogen and thereby produce the most effective immune response, and transfers much of the onus of self-non-self discrimination from the adaptive to the innate immune system. In the present article we highlight two paradoxes raised by the new model. First, by linking adaptive immunity to innate recognition the immune system is unable to take full advantage of the genetic ersity of T and B cell antigen receptors. Thus, the ability of the immune system to combat a pathogen is totally dependent on the efficiency of recognition by the innate system and, therefore, the germ-line mutation rate of the genes involved in the innate response. Second, if signals from the innate system induce costimulatory molecules on APC, then one would expect the accidental clonal expansion of many autoreactive T and B cells. We suggest that one means of resolving the first paradox is to propose that the major reason for the evolution of adaptive immunity was to provide, via immunological memory, resistance to reinfection, rather than simply to combat the primary infection by the pathogen. In the case of autoreactivity we suggest that autodestruction is prevented by immune responses being tightly regulated at the effector T cell level. Finally, we argue that the two paradoxes, rather than undermining the new model of immunity, highlight our lack of understanding of key elements of the vertebrate immune system.

Continual Low-Level Activation of the Classical Complement Pathway

Publisher: Rockefeller University Press

Date: 10-09-2001

DOI: 10.1084/JEM.194.6.747

Abstract: There is evidence that the classical complement pathway may be activated via a “C1-tickover” mechanism, analogous to the C3-tickover of the alternative pathway. We have quantitated and characterized this pathway of complement activation. Analysis of freshly collected mouse and human plasma revealed that spontaneous C3 activation rapidly occurred with the generation of C3 fragments in the plasma. By the use of complement- and Ig-deficient mice it was found that C1q, C4, C2, and plasma Ig were all required for this spontaneous C3 activation, with the alternative complement pathway further lifying C3 fragment generation. Study of plasma from a human with C1q deficiency before and after therapeutic C1q infusion confirmed the existence of a similar pathway for complement activation in humans. Elevated levels of plasma C3 were detected in mice deficient in complement components required for activation of either the classical or alternative complement pathways, supporting the hypothesis that there is continuous complement activation and C3 consumption through both these pathways in vivo. Blood stasis was found to stimulate C3 activation by classical pathway tick-over. This antigen-independent mechanism for classical pathway activation may augment activation of the complement system at sites of inflammation and infarction.

Extracellular histones induce erythrocyte fragility and anemia

Publisher: American Society of Hematology

Date: 28-12-2017

DOI: 10.1182/BLOOD-2017-06-790519

Abstract: Histones promote in vitro erythrocyte aggregation, sedimentation, fragility, and spleen retention in a concentration-dependent manner. Histones induce in vivo anemia, an increase in splenic hemoglobin content, as well as thrombocytopenia and leukopenia within a few minutes.

pDNA-lipoplexes engrafted with flagellin-related peptide induce potent immunity and anti-tumour effects

Publisher: Elsevier BV

Date: 09-2011

DOI: 10.1016/J.VACCINE.2011.07.045

Abstract: Complexes of cationic lipids and DNA (lipoplexes) are widely used for non-viral gene delivery and DNA vaccine development, but cationic lipids are toxic and promote non-specific interactions with cells, leading to poor efficacy. Near-neutral lipoplexes, on the other hand, can obviate toxicity, but a convenient means to target them to specific cells such as dendritic cells (DCs) has been lacking. Here, we show that a His-tagged flagellin-derived peptide (denoted 9Flg), previously reported to promote binding of liposomal antigen to TLR5-expressing cells, can be used to target near-neutral pDNA-lipoplexes incorporating the chelator lipid NTA(3)-DTDA (3(nitrilotriacetic acid)-ditetradecylamine) to DCs and other antigen-presenting cells (APCs). Thus, we show that pDNA-lipoplexes engrafted with 9Flg target pDNA to APCs in vitro and in vivo. Following i.v. administration, radiolabelled 9Flg-lipoplexes exhibited increased accumulation in spleen, lung and liver. Vaccination of C57BL/6 mice with 9Flg-lipoplexes containing either pcDNA3.1-SIIN (pSIIN) or a Kunjin virus replicon-based vector (pKUN), each encoding the epitope OVA(257-264) (SIINFEKL), induced Ag-specific T cell priming, and elicited strong cellular immunity as reflected by a marked increase in the number of Ag-responsive IFN-γ-producing CD8(+) T cells. Importantly, compared to i.m. injection of these SIINFEKL-encoding pDNAs in naked form, the i.v. administration of pSIIN or pKUN in 9Flg-lipoplexes to C57BL/6 mice induced a significantly more potent anti-tumour response in the B16-OVA melanoma tumour model. The targeting of near-neutral 9Flg-lipoplexes bearing pDNA encoding tumour antigens to TLR5 on APCs, therefore, is a powerful approach for developing more effective DNA vaccines and immunotherapies.

Th2-mediated anti-tumour immunity: friend or foe?

Publisher: Wiley

Date: 07-2007

DOI: 10.1111/J.1399-0039.2007.00869.X

Abstract: The concept that the immune system can recognise tumour cells and either eliminate them (tumour immune surveillance) or select for immunologically resistant variants (immunoediting) is gaining general acceptance by immunologists. In terms of an adaptive immune response to cancer, however, much of the research has focused on the response of cytotoxic CD8+ T lymphocytes to tumour-specific antigens and the production of Th1 cytokines by CD4+ and CD8+ T cells. In contrast, Th2-mediated immunity has traditionally been viewed as favouring tumour growth, both by promoting angiogenesis and by inhibiting cell-mediated immunity and subsequent tumour cell killing. While there is evidence that components of type 2 inflammation, such as B cells and interleukin-10, do promote tumour growth, there are also many studies demonstrating the anti-tumour activity of CD4+ Th2 cells, particularly in collaboration with tumour-infiltrating granulocytes, such as eosinophils. In this review, we examine all the components of type 2 immunity and their effects on tumour growth. Collectively, from this analysis, we conclude that there is a great potential for the development of Th2-mediated immunotherapies that harness the cytotoxic activity of eosinophils.

Tolerance in Adult Rats to a Purified Protein, Flagellin, from Salmonella adelaide

Publisher: Springer Science and Business Media LLC

Date: 09-1967

DOI: 10.1038/2151202A0

Abstract: The main purpose of the present study was to assess the impact of global positioning system (GPS) signal lapse on physical activity analyses, discover any existing associations between missing GPS data and environmental and demographics attributes, and to determine whether imputation is an accurate and viable method for correcting GPS data loss. Accelerometer and GPS data of 782 participants from 8 studies were pooled to represent a range of lifestyles and interactions with the built environment. Periods of GPS signal lapse were identified and extracted. Generalised linear mixed models were run with the number of lapses and the length of lapses as outcomes. The signal lapses were imputed using a simple ruleset, and imputation was validated against person-worn camera imagery. A final generalised linear mixed model was used to identify the difference between the amount of GPS minutes pre- and post-imputation for the activity categories of sedentary, light, and moderate-to-vigorous physical activity. Over 17% of the dataset was comprised of GPS data lapses. No strong associations were found between increasing lapse length and number of lapses and the demographic and built environment variables. A significant difference was found between the pre- and postimputation minutes for each activity category. No demographic or environmental bias was found for length or number of lapses, but imputation of GPS data may make a significant difference for inclusion of physical activity data that occurred during a lapse. Imputing GPS data lapses is a viable technique for returning spatial context to accelerometer data and improving the completeness of the dataset.

Regulation of the immune response. I. Suppression of delayed-type hypersensitivity by T cells from mice expressing humoral immunity

Publisher: Wiley

Date: 10-1976

DOI: 10.1002/EJI.1830061003

Abstract: The ability of horse red blood cell (HRBC)-specific T cells from mice expressing humoral immunity to suppress the induction of HRBC-specific delayed-type hypersensitivity (DTH) was investigated. The transfer of Ig-negative spleen cells, from mice injected 4 days previously with HRBC, completely suppressed the development of DTH in mice treated with cyclophosphamide and sensitized with HRBC. The suppressor cell was found to be lysed by treatment with anti-theta serum and complement. Furthermore, hemocyanin-specific immune T cells were able to suppress the DTH induced to HRBC, provided these two antigens were coupled together. These studies suggest that T cells present under conditions were humoral immunity is induced can suppress DTH and that such cells play an important role in the regulation of the immune response.

Plasmodium yoelii: antibody and the maintenance of immunity in BALB/c mice.

Publisher: Elsevier BV

Date: 08-1981

DOI: 10.1016/0014-4894(81)90056-4

Islet heparan sulfate but not heparan sulfate proteoglycan core protein is lost during islet isolation and undergoes recovery post-islet transplantation

Publisher: Elsevier BV

Date: 11-2015

DOI: 10.1111/AJT.13366

Abstract: Islet beta cells in situ express intracellular heparan sulfate (HS), a property previously shown in vitro to be important for their survival. We report that HS levels inside islet beta cells correlate with the novel intracellular localization of the HSPG core proteins for collagen type XVIII (Col18), a conventional extracellular matrix component. Syndecan-1 (Sdc1) and CD44 core proteins were similarly localized inside beta cells. During isolation, mouse islets selectively lose HS to 11-27% of normal levels but retain their HSPG core proteins. Intra-islet HS failed to recover substantially during culture for 4 days and was not reconstituted in vitro using HS mimetics. In contrast, significant recovery of intra-islet HS to ∼40-50% of normal levels occurred by 5-10 days after isotransplantation. Loss of islet HS during the isolation procedure is independent of heparanase (a HS-degrading endoglycosidase) and due, in part, to oxidative damage. Treatment with antioxidants reduced islet cell death by ∼60% and increased the HS content of isolated islets by ∼twofold compared to untreated islets, preserving intra-islet HS to ∼60% of the normal HS content of islets in situ. These findings suggest that the preservation of islet HS during the islet isolation process may optimize islet survival posttransplant.

Plasminogen Is Tethered with High Affinity to the Cell Surface by the Plasma Protein, Histidine-rich Glycoprotein

Publisher: Elsevier BV

Date: 09-2004

DOI: 10.1074/JBC.M406027200

Topographical studies of lymphocyte localization using an intracellular fluorochrome

Publisher: Wiley

Date: 11-1985

DOI: 10.1002/AR.1092130308

Abstract: A procedure for analysing the topographical localization in tissue sections or whole-organ mounts of lymphocytes labelled with an intracellular DNA-binding fluorochrome, Hoechst dye No. 33342, is described. The localization of intravenously injected lymphocytes in spleen, popliteal lymph nodes, and Peyer's patches was followed up to 7 days. In the case of spleen, both B and T lymphocytes initially localised in the marginal zone. Subsequently, B cells appeared to exit via the red pulp, while T cells aggregated around vessels in the white pulp. In Peyer's patches, B and T lymphocytes localized to different lymphoid areas. The advantages and potential applications of this technique are discussed.

Monitoring lymphocyte proliferation in vitro and in vivo with the intracellular fluorescent dye carboxyfluorescein diacetate succinimidyl ester

Publisher: Springer Science and Business Media LLC

Date: 23-08-2007

DOI: 10.1038/NPROT.2007.296

Abstract: This protocol outlines the carboxyfluorescein diacetate succinimidyl ester (CFSE) method for following the proliferation of human lymphocytes in vitro and mouse lymphocytes both in vitro and in vivo. The method relies on the ability of CFSE to covalently label long-lived intracellular molecules with the highly fluorescent dye, carboxyfluorescein. Following each cell ision, the equal distribution of these fluorescent molecules to progeny cells results in a halving of the fluorescence of daughter cells. The CFSE labeling protocol described, which typically takes <1 h to perform, allows the detection of up to eight cell isions before CFSE fluorescence is decreased to the background fluorescence of unlabeled cells. Protocols are outlined for labeling large and small numbers of human and mouse lymphocytes, labeling conditions being identified that minimize CFSE toxicity but maximize the number of cell isions detected. An important feature of the technique is that ision-dependent changes in the expression of cell-surface markers and intracellular proteins are easily quantified by flow cytometry.

Pancreatic Islet Basement Membrane Loss and Remodeling after Mouse Islet Isolation and Transplantation: Impact for Allograft Rejection

Publisher: SAGE Publications

Date: 2014

DOI: 10.3727/096368912X659880

Abstract: The isolation of islets by collagenase digestion can cause damage and impact the efficiency of islet engraftment and function. In this study, we assessed the basement membranes (BMs) of mouse pancreatic islets as a molecular biomarker for islet integrity, damage after isolation, and islet repair in vitro as well as in the absence or presence of an immune response after transplantation. Immunofluorescence staining of BM matrix proteins and the endothelial cell marker platelet endothelial cell adhesion molecule-1 (PECAM-1) was performed on pancreatic islets in situ, isolated islets, islets cultured for 4 days, and islet grafts at 3–10 days posttransplantation. Flow cytometry was used to investigate the expression of BM matrix proteins in isolated islet β-cells. The islet BM, consisting of collagen type IV and components of Engelbreth–Holm–Swarm (EHS) tumor laminin 111, laminin α2, nidogen-2, and perlecan in pancreatic islets in situ, was completely lost during islet isolation. It was not reestablished during culture for 4 days. Peri- and intraislet BM restoration was identified after islet isotransplantation and coincided with the migration pattern of PECAM-1 + vascular endothelial cells (VECs). After islet allotransplantation, the restoration of VEC-derived peri-islet BMs was initiated but did not lead to the formation of the intraislet vasculature. Instead, an abnormally enlarged peri-islet vasculature developed, coinciding with islet allograft rejection. The islet BM is a sensitive biomarker of islet damage resulting from enzymatic isolation and of islet repair after transplantation. After transplantation, remodeling of both peri- and intraislet BMs restores β-cell–matrix attachment, a recognized requirement for β-cell survival, for isografts but not for allografts. Preventing isolation-induced islet BM damage would be expected to preserve the intrinsic barrier function of islet BMs, thereby influencing both the effector mechanisms required for allograft rejection and the antirejection strategies needed for allograft survival.

Venular and Arterial Endothelial Cells Differ in their Expression of Adhesion Molecules and in their Ability to Degrade the Subendothelial Basement Membrane

Publisher: Informa UK Limited

Date: 1995

DOI: 10.3109/10623329509024650

Heparan sulfate: A ubiquitous glycosaminoglycan with multiple roles in immunity

Publisher: Frontiers Media SA

Date: 2013

DOI: 10.3389/FIMMU.2013.00470

Promotion of mammalian angiogenesis by neolignans derived from soybean extracellular fluids

Publisher: Public Library of Science (PLoS)

Date: 08-05-2018

DOI: 10.1371/JOURNAL.PONE.0196843

Heparanase and autoimmune diabetes

Publisher: Frontiers Media SA

Date: 2013

DOI: 10.3389/FIMMU.2013.00471

Circulating platelet-neutrophil aggregates characterize the development of type 1 diabetes in humans and NOD mice

Publisher: American Society for Clinical Investigation

Date: 25-01-2022

DOI: 10.1172/JCI.INSIGHT.153993

Polysaccharides influence the aggregation of Dictyostelium discoideum cells and bind to developmentally regulated cell surface proteins

Publisher: Elsevier BV

Date: 04-1993

DOI: 10.1006/EXCR.1993.1100

Abstract: Six of ten anionic polysaccharides studied were found to significantly reduce the adhesion of growth-phase Dictyostelium discoideum cells. However, only hyaluronic acid, chondroitin-4-sulfate and chondroitin-6-sulfate interfered with the adhesion of aggregation-competent cells. Neither EDTA-stable nor EDTA-sensitive adhesion of postaggregation cells were affected by the polyanions. The two chondroitin sulfates influenced the aggregation of cells in submerged cultures, long and broad aggregation streams being formed and the broad sheets of cells eventually building multilayered aggregates. Radioiodination of cell surface proteins followed by cellulose fiber affinity chromatography identified the same nine proteins bound by hyaluronic acid and the chondroitin sulfates, six of which were regulated during development. Protease-resistant anionic material isolated from cells bound the same surface proteins as the three glycosaminoglycans. Discoidin I bound to the uncoupled cellulose fibers, suggesting a structural role for the lectin in the extracellular slime sheath. Anionic polysaccharides and cell surface lectins that bind them may be involved in the cell recognition, cell aggregation, and the cell sorting that occurs during pattern formation.

Evidence for cell surface association of CD2 and LFA‐1 (CD11a/CD18) on T lymphocytes

Publisher: Wiley

Date: 02-1994

DOI: 10.1002/EJI.1830240228

Abstract: Previous studies have reported an association of the cell surface adhesion molecule CD2 with the T cell receptor and with CD45 on mouse and human T lymphocytes. In this study the association of CD2 with cell surface molecules was investigated using cell surface biotinylation of T lymphocytes, coupled with immunoprecipitation using two CD2-specific monoclonal antibodies (mAb) (RM2-5 and 12-15) and analysis by SDS-PAGE. Although both CD2 mAb immunoprecipitated CD2 from lysates of murine lymphocytes, it was found that mAb 12-15, but not RM2-5, co-precipitated two other molecules of 95 and 180 kDa. Subsequent studies revealed that the 95- and 180-kDa molecules were associated with a subspecies of CD2 (approximately 5%) on thymocytes, the antigen-specific T cell line D10, and splenic T cells but not B cells. Two lines of evidence were obtained consistent with the 95- and 180-kDa molecules being the beta and alpha chains of LFA-1. Firstly, an analysis of 12-15 mAb immunoprecipitates on 4-12% gels under reducing and nonreducing conditions shows that the 95- and 180-kDa molecules have a molecular weight and migration pattern identical to LFA-1. Secondly, depletion of LFA-1 from lysates with LFA-1 mAb abolished the ability of CD2 mAb 12-15 to co-precipitate the 95- and 180-kDa molecules, thereby identifying these as the beta and alpha chains of mouse LFA-1, respectively. These results provide evidence for the first time for an association of LFA-1 and CD2 on mouse T lymphocytes, and suggest that the association occurs with an immunologically distinct subspecies of CD2 molecules.

Evidence that Thy-1 and Ly-5 (T-200) antigens interact with sulphated carbohydrates

Publisher: Wiley

Date: 06-1988

DOI: 10.1038/ICB.1988.28

Abstract: Recent studies have demonstrated that lymphocytes express an array of cell surface receptors for sulphated polysaccharides (SP). Experiments were undertaken to determine the binding characteristics of these receptors and establish whether any known lymphocyte cell surface antigens interact with sulphated carbohydrates. It was found that murine thymocytes lack receptors for chondroitin-4-sulphate but express saturable, high affinity binding sites for heparin, fucoidan and dextran sulphate, with an apparent affinity constant range of 0.03-2.6 x 10(-9) mol/l. Binding inhibition experiments revealed one class of binding sites on murine thymocytes that is shared by heparin, fucoidan and dextran sulphate and another class of sites that is dextran sulphate-specific. The cell surface receptors for the SP were affinity-purified by applying detergent lysates of 125I-labelled thymocyte membranes to SP-coupled solid supports. It was found that the Thy-1 and Ly-5 (T-200 or leucocyte common antigen) molecules of murine thymocytes bind to sulphated carbohydrates, although the two molecules differed substantially in their reactivity with the four different SP tested. Furthermore, only subpopulations of the Thy-1 and Ly-5 molecules interacted with sulphated sugars. Four additional sulphated carbohydrate-binding molecules were also detected. It is suggested that the SP-binding molecules are involved in the interaction of lymphocytes with glycosaminoglycans on other cells and in the interstitial space.

Evaluation of the ability of digital infrared imaging to detect vascular changes in experimental animal tumours

Publisher: Wiley

Date: 2004

DOI: 10.1002/IJC.11618

Abstract: Infrared imaging has frequently been used in the past to detect changes in skin surface temperature associated with breast cancer. Usually a 1-2 degrees C elevation in skin surface temperature is observed at the tumour periphery, and it has been proposed that this change is due to hypervascularity resulting from tumour-associated angiogenesis. In our study, we used the rat mammary adenocarcinoma 13762 MAT, a tumour that has been used to identify antiangiogenic drugs, to investigate whether infrared imaging can detect angiogenesis in malignant tumours. If successful, it was hoped that this technique would represent a simple, noninvasive, procedure for monitoring the activity of antiangiogenic drugs. It was found that, unlike breast cancer patients, no tumour-associated increase in skin surface temperature was observed, but a constant and highly significant reduction in temperature was noted that was independent of tumour size and was produced by relatively small tumours (>/= 0.5 cm in diameter). The explanation for this effect is unclear but it may be due to the poorly vascularised nature of rapidly growing tumours. Nevertheless, our study indicates that the peripheral temperature elevation reported in breast cancer patients is unlikely to be due to hypervascularity resulting from tumour-induced angiogenesis. An alternative explanation is that the temperature increase is due to a chronic inflammatory response around developing breast tumours. With increasing evidence that inflammation can enhance tumour growth and is associated with a poor prognosis, this suggestion implies that infrared imaging may have considerable prognostic value.

The use of CFSE-like dyes for measuring lymphocyte proliferation: Experimental considerations and biological variables

Publisher: EDP Sciences

Date: 2012

DOI: 10.1051/MMNP/20127505

Destroying c-jun messenger: New insights into biological mechanisms of DNAzyme Function

Publisher: Impact Journals, LLC

Date: 30-06-2012

DOI: 10.18632/ONCOTARGET.549

Separation and Functional Analysis of Subpopulations of Lymphocytes Bearing Complement and Fc Receptors

Publisher: Wiley

Date: 08-1975

DOI: 10.1111/J.1600-065X.1975.TB00727.X

Abstract: A highly versatile procedure is described in this review which can be used to separate and obtain in pure form subpopulations of lymphoid cells which express different cell surface structures. The method is based on the observation that when rosetting and non-rosetting leukocytes are centrifuged on a cushion of Isopaque/Ficoll, the rosetting leukocytes and red cells sink whereas the non-rosetting leukocytes float. Thus, any subpopulation of leukocytes can be separated providing they can be identified by rosetting. The earlier sections of this review describe the method, its efficiency of separation and its advantages compared with other fractionation procedures. Subsequent sections describe experiments in which the procedure was specifically applied to separating Fc receptor (Fc+) and complement receptor (CR+) lymphocytes. On the basis of these two receptors it was possible to sub ide T and B lymphocytes into distinct subpopulations. Four subclasses of B lymphocytes were identified in mouse spleen (Fc+CR+,Fc+CR-,Fc-CR+ and Fc-CR-) and two subclasses of T cells were also detected (Fc+ and Fc-). The functional relevance of these subpopulations of lymphocytes was examined. It was found that in all cases examined, antigens could successfully activate CR+ B cells to produce antibody. However, only polymeric antigens, whether T-dependent or T-independent, were capable of triggering CR- B cells to synthesize antibody. Furthermore, preliminary experiments suggest that Fc receptors are present on functional B cells and helper T cells but are not expressed on cytotoxic T cells. On the basis of these results it is proposed that complement receptors on B lymphocytes provide an additional binding site which stabilizes the union between the antigen-specific receptors and soluble antigen. In contrast, due to their multi-determinant nature, polymeric antigens can avidly bind to B cells without involvement of the complement receptors. The possibility of Fc receptors playing a similar role in stabilizing the interaction of antigen with specific receptors on lymphocytes, particularly on T helper cells, is also discussed.

Targeted therapies in the management of locally advanced and metastatic pancreatic cancer: A systematic review

Publisher: Impact Journals, LLC

Date: 20-04-2018

DOI: 10.18632/ONCOTARGET.25085

Evidence that mannose recognition by splenic sinusoidal cells plays a role in the splenic entry of lymphocytes

Publisher: Wiley

Date: 08-1992

DOI: 10.1002/EJI.1830220804

Abstract: Previous studies demonstrated that mannan is a potent inhibitor of splenic entry of lymphocytes and mediates its inhibitory effect at an unidentified site in the spleen rather than acting directly on lymphocytes. This report describes the in vivo site of action of mannan. In vivo localization studies with fluoresceinated preparations of mannan (Fl-mannan) and a mannose-6-phosphate-containing yeast phosphomannan monoester core from P. holstii exopolysaccharide (Fl-PPME) demonstrated that the polysaccharide specifically localize in the splenic marginal sinuses in cells with a dendritic morphology termed splenic sinusoidal cells (SSC). Uptake of the polysaccharides by SSC was mediated by a mannan-specific receptor which was saturable and of high avidity. Several lines of evidence suggested that mannan uptake by SSC inhibited splenic entry of lymphocytes. First, the ability of SSC to bind Fl-mannan and Fl-PPME closely paralleled the ability of these polysaccharides to inhibit splenic entry of lymphocytes. In fact, doses of mannan and PPME which would saturate SSC mannan receptors completely blocked splenic entry of lymphocytes. Second, SSC are situated at the initial entry point of lymphocytes into spleen and passage of lymphocytes through the SSC region of spleen was profoundly inhibited by mannan. Finally, direct evidence for adhesion between lymphocytes and SSC was obtained with spleen cell suspensions where clustering between Fl-mannan labeled SSC and lymphocytes was observed. Collectively, these data indicate that mannan (and PPME) inhibit splenic entry of lymphocytes by interacting with SSC, cell which play a critical role in the entry of lymphocytes into the spleen. Whether mannan-specific receptors on SSC directly mediate lymphocyte-SSC adhesion or play on indirect role in modifying lymphocyte migration requires further investigation.

The effect of sulfated polysaccharides on the free intracellular calcium ion concentration of lymphocytes

Publisher: Elsevier BV

Date: 19-08-1987

DOI: 10.1016/0167-4889(87)90155-8

Abstract: Recent studies have demonstrated that murine lymphocytes express specific cell-surface receptors for a range of sulfated polysaccharides. In order to determine whether polysaccharide binding induces transmembrane signaling, the effects of sulfated polysaccharides on the free intracellular calcium ion concentration [( Ca2+]i) of mouse thymocytes and spleen cells were determined. Cells were loaded with Indo-I, a fluorescent indicator of calcium ion concentration. The validity and limitations in the use of this indicator in the determination of [Ca2+]i are documented. Dextran sulfate (Mn = 500,000), iota-carrageenan, lambda-carrageenan and kappa-carrageenan all cause relatively large changes in the [Ca2+]i of thymocytes (change in [Ca2+]i greater than 50 nM). Of these, dextran sulfate (Mn = 500,000) always had the greatest effect on [Ca2+]i. Smaller responses were obtained with heparin and dextran sulfate (Mn = 5000), while no response was obtained with chondroitin 4-sulfate, chondroitin 6-sulfate, pentosan sulfate or fucoidin. This response pattern (with the exception of fucoidin and pentosan sulfate) corresponds with the expression of thymocyte receptors for these polysaccharides. The increase in [Ca2+]i caused by the sulfated polysaccharides requires extracellular Ca2+ ions however, it is unlikely that voltage-dependent ion channels are involved in these responses. In contrast to thymocytes, although spleen cells express receptors for sulfated polysaccharides, they were unresponsive to all of the sulfated polysaccharides tested, suggesting a basic difference between thymocytes and peripheral T and B lymphocytes in their response to the binding of sulfated polysaccharides.

Involvement of Cells With ‘B’ Properties in Development of T Helpers

Publisher: Springer US

Date: 1976

DOI: 10.1007/978-1-4613-4355-4_10

Abstract: Cells bearing Ig on the membrane, fractionated from either 17 day embryonic livers or from normal adult bone marrow, when transferred to splenectomized-irradiated mice, lead to development of helper cells in the thymus of the recipients. The helper function was expressed when the recipients were stimulated with flagellin-MON and the thymus cells were cultured together with anti theta treated spleen cells and stimulated with DNP-MON. The response to DNP was not enhanced when the irradiated mice were inoculated with non Ig bearing cells. Helper activity was related to cells which were eliminated with anti theta antibodies and did not have detectable Ig. Hence, cells with 'B' properties may be involved in development of T helpers.

Automated rosetting assay for cell surface antigens and autorosetting lymphocytes

Publisher: Elsevier BV

Date: 12-1982

DOI: 10.1016/0022-1759(82)90093-X

Abstract: An automated procedure, using a Coulter counter, is described for enumerating rosette-forming lymphocytes in 2 rosetting systems in mice, detecting antibodies to cell surface antigens, and the interaction of autologous erythrocytes with thymocytes (autorosetting). The procedure gives results comparable with determinations of rosettes by light microscopy. The procedure not only estimates rosetting percentages, but can be used to titrate anti-lymphocyte antibodies, to detect autorosette inhibition factor in serum and to assay cell surface antigens in detergent lysates of spleen cells.

The Tissue Localization, Immunogenic, and Tolerance-Inducing Properties of Antigens and Antigen-Fragments

Publisher: Cold Spring Harbor Laboratory

Date: 1967

DOI: 10.1101/SQB.1967.032.01.050

Molecular composition of the peri-islet basement membrane in NOD mice: A barrier against destructive insulitis

Publisher: Springer Science and Business Media LLC

Date: 17-07-2008

DOI: 10.1007/S00125-008-1085-X

Cleavage of bacterial flagellin with cyanogen bromide. Chemical and physical properties of the protein fragments

Publisher: Portland Press Ltd.

Date: 07-1969

DOI: 10.1042/BJ1130489

Abstract: 1. Flagellin, isolated from the flagella of Salmonella adelaide, was shown by various criteria to be a pure protein. It had a molecular weight of about 40000 and contained three methionine, six tyrosine, 11 arginine and 25 lysine residues/mol., of which 11 of the lysine residues were present as ∈-N-methyl-lysine. 2. After treatment of flagellin with cyanogen bromide in formic acid, four main fragments (A, B, C and D) were obtained, with as many as six minor components that represented partial degradation products. The major fragments were estimated by amino acid analysis to have molecular weights of about 18000 for fragment A, 12000 for fragment B, 5500 for fragment C and 4500 for fragment D. Fragments A, B and D, but not fragment C, were recovered pure by gel chromatography as monitored by polyacrylamide-gel electrophoresis. 3. A complex between fragments C and D was also isolated (mol.wt. 10000) after limited oxidation of flagellin by chloramine-t before digestion by cyanogen bromide. After oxidation essentially only two fragments were released from flagellin by cyanogen bromide: the ‘C,D’ complex and a presumed ‘AB’ fragment. 4. The sum of the amino acid analyses of fragments A and B and the ‘C,D’ complex gave residue values that agreed well with the amino acid composition of native flagellin. 5. Fragments A and D contained tyrosine, and ten of the 11 ∈-N-methyl-lysine residues of the molecule were in fragment A. Reaction with [125I]iodide at small extents of substitution showed that, in flagellin, the tyrosine residue of fragment D was more readily substituted than those of fragment A. By contrast, in polymerized flagellin, the tyrosine residues of fragment A were more readily substituted. 6. Treatment of flagellin with carboxypeptidases A and B revealed the C-terminal sequence -Leu-Leu-Leu-Arg. Arginine and leucine were released by carboxypeptidase from the ‘C,D’ complex but not from fragment D, indicating that fragment C was C-terminal. 7. On the basis of the results from amino acid analysis, carboxypeptidase digestion, N-terminal analysis, iodination studies and polyacrylamide-gel electrophoresis, the sequence of fragments in flagellin was considered to be B–A–D–C in the polymer, fragment A was exposed. It is suggested that methylation of the lysine residues occurred in the organism after flagellin had polymerized.

Immune response to chemically modified flagellin. II. Evidence for a fundamental relationship between humoral and cell-mediated immunity

Publisher: Rockefeller University Press

Date: 07-1971

DOI: 10.1084/JEM.134.1.21

Abstract: Flagellin (mol.wt. 40,000) from S. adelaide organisms and a series of acetoacetyl derivatives of flagellin were tested for their ability to induce humoral and cell-mediated immunity in adult rats. It was found that unmodified flagellin was an excellent inducer of antibody formation but a poor inducer of delayed-type hypersensitivity. In contrast, increasing acetoacetylation steadily destroyed the ability of flagellin to initiate antibody formation but enhanced the capacity of the molecule to induce flagellin-specific cell-mediated immunity and antibody tolerance. In fact, it appeared that in adult rats antibody formation and cell-mediated immunity may well be opposing immunological processes. Furthermore, the affinity of the acetoacetyl flagellins for anti-flagellin antibodies appeared to determine the type of immune response which predominated. High affinity antigen produced antibody formation whereas low affinity antigen induced cell-mediated immunity and antibody tolerance. The importance of affinity was further evidenced by the fact that a CNBr digest of flagellin induced humoral and cellular immune responses identical to an acetoacetylated flagellin of comparable antigenic activity. From these studies it was proposed that both humoral and cell-mediated immunity can be directed against the same antigenic determinants but that the specificity requirements for delayed hypersensitivity (and antibody tolerance) are less than those required for antibody formation. Some remarkable immunological features of the flagellin system were revealed. Flagellin induced comparable delayed-type hypersensitivity when injected in either saline or FCA. Furthermore, FCA only slightly enhanced the delayed responses induced by the acetoacetyl flagellins and in fact these preparations produced antibody tolerance whether injected in saline or adjuvant. Finally, in contrast to the adult tolerance induced by the acetoacetylated flagellins, which existed only at the antibody level, tolerance in neonatal rats existed at the level of both humoral and cell-mediated immunity. This finding is the first indication of a fundamental difference between neonatal and adult tolerance. The significance of these findings is discussed in the light of current immunological concepts and a hypothesis proposed to explain these phenomena.

Histidine-Rich Glycoprotein and Platelet Factor 4 Mask Heparan Sulfate Proteoglycans Recognized by Acidic and Basic Fibroblast Growth Factor

Publisher: American Chemical Society (ACS)

Date: 11-1994

DOI: 10.1021/BI00250A047

Abstract: Recent studies have shown that fibroblast growth factors (FGFs) need to interact with cell-surface heparan sulfate proteoglycans (HSPGs) in order to bind to and activate FGF receptors. In this paper, three major heparin-binding proteins, histidine-rich glycoprotein (HRG) and antithrombin III (ATIII), which are constitutively present at high concentrations in plasma, and platelet factor 4 (PF4), which is released locally at high concentrations by degranulating platelets, were tested for their ability to act as modulators of FGF activity by competing with the FGFs for cell-surface HSPGs. HRGs from both chicken and human, and human PF4, were demonstrated to compete with each other and with acidic FGF (aFGF) and basic FGF (bFGF) for binding to BALB/c 3T3 cell-surface HSPGs, whereas ATIII did not compete. Thus, HRG, PF4, aFGF, and bFGF all interact with the same HS chains on the 3T3 cell surface, either binding to the same or binding to adjacent saccharide sequences on the chains. In terms of their relative binding affinity for cell-surface HSPGs, the hierarchy was shown to be PF4 > or = bFGF > aFGF = cHRG > hHRG. HRG was also shown to significantly inhibit both FGF-stimulated and endogenous 3T3 cell DNA synthesis. HRG also binds to extracellular matrices (ECM), originating from bovine corneal endothelial cells, in a heparin-inhibitable manner. Indeed, both HRG and PF4, at physiological concentrations, were shown to effectively inhibit the binding of 125I-aFGF and 125I-bFGF to ECM. In addition, HRG was able to displace biologically active bFGF from the ECM. On the basis of these findings, it is proposed that HRG and PF4 may act as positive regulators of FGF activity by displacing FGF from the ECM or basement membrane and making FGF available to responsive cells. Alternatively, they could act as negative regulators by masking HSPGs on responsive cells and preventing FGF receptor activation.

Anti-self receptors. IV. H-2-restricted receptors on thymocytes recognize carbohydrate structures on target cells.

Publisher: Springer Science and Business Media LLC

Date: 12-1981

DOI: 10.1007/BF01561699

Abstract: An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Discrimination of suppressor T cells of humoral and cell-mediated immunity by anti-Ly and anti-Ia sera

Publisher: Elsevier BV

Date: 06-1977

DOI: 10.1016/0008-8749(77)90038-7

Abstract: The cellular uptake of N-acetyl-alpha-D-glucosaminidase, the deficient enzyme in Sanfilippo B disease, and the intracellular fate and metabolic effect of this enzyme have been investigated in Sanfilippo B and normal fibroblasts. For both genotypes the uptake is highly efficient (up to 0.025 mU/h/mg cell protein), specific and constant over a period of at least 6 days. It is probable that the enzyme protein is taken up by adsorptive pinocytosis. The enzymatic activity as well as the biological activity towards (35)SO(4)-labeled mucopolysaccharides persist in Sanfilippo B cells with a half-life of 34 h, indicating the intralysosomal localization of the pinocytosed enzyme. The data obtained are discussed with regard to a possible enzyme replacement therapy. For Sanfilippo B disease the doses used in the past are considered to be insufficient to cause measurable effects.

Fluorescent target array killing assay: A multiplex cytotoxic T-cell assay to measure detailed T-cell antigen specificity and avidity in vivo

Publisher: Wiley

Date: 27-06-2012

DOI: 10.1002/CYTO.A.22084

Abstract: Here we describe a multiplex, fluorescence-based, in vivo cytotoxic T-cell assay using the three vital dyes carboxyfluorescein diacetate succinimidyl ester, cell trace violet, and cell proliferation dye efluor 670. When used to label cells in combination, these dyes can discriminate >200 different target cell populations in the one animal due to each target population having a unique fluorescence signature based on fluorescence intensity and the different emission wavelengths of the dyes. This allows the simultaneous measurement of the in vivo killing of target cells pulsed with numerous peptides at different concentrations and the inclusion of many replicates. This fluorescent target array killing assay can be used to measure the fine antigen specificity and avidity of polyclonal cytotoxic T-cell responses in vivo, immunological parameters that were previously impossible to monitor.

Neutrophil extracellular traps and their histones promote Th17 cell differentiation directly via TLR2

Publisher: Springer Science and Business Media LLC

Date: 26-01-2022

DOI: 10.1038/S41467-022-28172-4

Abstract: Neutrophils perform critical functions in the innate response to infection, including through the production of neutrophil extracellular traps (NETs) - web-like DNA structures which are extruded from neutrophils upon activation. Elevated levels of NETs have been linked to autoimmunity but this association is poorly understood. By contrast, IL-17 producing Th17 cells are a key player in various autoimmune diseases but are also crucial for immunity against fungal and bacterial infections. Here we show that NETs, through their protein component histones, directly activate T cells and specifically enhance Th17 cell differentiation. This modulatory role of neutrophils, NETs and their histones is mediated downstream of TLR2 in T cells, resulting in phosphorylation of STAT3. The innate stimulation of a specific adaptive immune cell subset provides an additional mechanism demonstrating a direct link between neutrophils, NETs and T cell autoimmunity.

Immune response to chemically modified flagellin. I. Induction of antibody tolerance to flagellin by acetoacetylated derivatives of the protein

Publisher: Rockefeller University Press

Date: 07-1971

DOI: 10.1084/JEM.134.1.1

Abstract: Flagellin (mol. wt. 40,000) from S. adelaide organisms was acetoacetylated to varying extents with diketene (acetoacetic anhydride). Chemical studies demonstrated that the amino groups of flagellin were more readily acetoacetylated than the hydroxyl groups. Several antigenic tests revealed that as flagellin was acetoacetylated to increasing extents there was a steady decline in the affinity of the molecule for anti-flagellin antibodies. Loss in antigenic activity following acetoacetylation was not related to the number of acetoacetyl groups attached but was determined by the type of residue substituted. Reactive lysine residues were much less important anti-genically than easily substituted hydroxyl groups. Acetoacetylation very readily destroyed the antibody-forming capacity of flagellin in rats. This fall in immunogenicity was related to the antigenic activity of the preparations. In fact, only a 40% reduction in the antigenic activity of flagellin produced a 90–95% reduction in primary antibody formation. The more heavily acetoacetylated flagellins produced no detectable antibody and, in fact, rendered adult rats tolerant (in terms of antibody formation) to a subsequent challenge of flagellin. Tolerance was induced by acetoacetylated flagellins which had drastically reduced affinities for anti-flagellin antibodies. These results were interpreted as indicating that the affinity of antigen for receptors on cells appears to be of crucial importance in determining whether antibody formation or immunological tolerance (antibody suppression) occurs.

Immunotherapy of cytotoxic T cell-resistant tumors by T helper 2 cells: An eotaxin and STAT6-dependent process

Publisher: Rockefeller University Press

Date: 27-01-2003

DOI: 10.1084/JEM.20021683

Abstract: Currently most attempts at cancer immunotherapy involve the generation of CD8+ cytotoxic T lymphocytes (CTLs) against tumor-associated antigens. Many tumors, however, have been immunoselected to evade recognition by CTLs and thus alternative approaches to cancer immunotherapy are urgently needed. Here we demonstrate that CD4+ T cells that recognize a secreted tumor-specific antigen and exhibit a cytokine secretion profile characteristic of Th2 cells, are capable of clearing established lung and visceral metastases of a CTL-resistant melanoma. Clearance of lung metastases by the Th2 cells was found to be totally dependent on the eosinophil chemokine, eotaxin, and partially dependent on the transcription activator signal transducer and activator of transcription 6 (STAT6), with degranulating eosinophils within the tumors inducing tumor regression. In contrast, tumor-specific CD4+ Th1 cells, that recruited macrophages into the tumors, had no effect on tumor growth. This work provides the basis for a new approach to adoptive T cell immunotherapy of cancer.

Human HPA endoglycosidase heparanase. Map position 4q21.3

Publisher: Springer Science and Business Media LLC

Date: 1999

DOI: 10.1023/A:1009235132576

Heparan sulfate proteoglycans in beta cells provide a critical link between endoplasmic reticulum stress, oxidative stress and type 2 diabetes

Publisher: Public Library of Science (PLoS)

Date: 04-06-2021

DOI: 10.1371/JOURNAL.PONE.0252607

Abstract: Heparan sulfate proteoglycans (HSPGs) consist of a core protein with side chains of the glycosaminoglycan heparan sulfate (HS). We have previously identified (i) the HSPGs syndecan-1 (SDC1), and collagen type XVIII (COL18) inside mouse and human islet beta cells, and (ii) a critical role for HS in beta cell survival and protection from reactive oxygen species (ROS). The objective of this study was to investigate whether endoplasmic reticulum (ER) stress contributes to oxidative stress and type 2 diabetes (T2D) by depleting beta cell HSPGs/HS. A rapid loss of intra-islet/beta cell HSPGs, HS and heparanase (HPSE, an HS-degrading enzyme) accompanied upregulation of islet ER stress gene expression in both young T2D-prone db/db and Akita Ins2 WT/C96Y mice. In MIN6 beta cells, HSPGs, HS and HPSE were reduced following treatment with pharmacological inducers of ER stress (thapsigargin or tunicamycin). Treatment of young db/db mice with Tauroursodeoxycholic acid (TUDCA), a chemical protein folding chaperone that relieves ER stress, improved glycemic control and increased intra-islet HSPG/HS. In vitro , HS replacement with heparin (a highly sulfated HS analogue) significantly increased the survival of wild-type and db/db beta cells and restored their resistance to hydrogen peroxide-induced death. We conclude that ER stress inhibits the synthesis/maturation of HSPG core proteins which are essential for HS assembly, thereby exacerbating oxidative stress and promoting beta cell failure. Diminished intracellular HSPGs/HS represent a previously unrecognized critical link bridging ER stress, oxidative stress and beta cell failure in T2D.

Reversal of the glycolytic phenotype by dichloroacetate inhibits metastatic breast cancer cell growth in vitro and in vivo

Publisher: Springer Science and Business Media LLC

Date: 19-06-2009

DOI: 10.1007/S10549-009-0435-9

Abstract: The glycolytic phenotype is a widespread phenomenon in solid cancer forms, including breast cancer. Dichloroacetate (DCA) has recently been proposed as a novel and relatively non-toxic anti-cancer agent that can reverse the glycolytic phenotype in cancer cells through the inhibition of pyruvate dehydrogenase kinase. We have examined the effect of DCA against breast cancer cells, including in a highly metastatic in vivo model. The growth of several breast cancer cell lines was found to be inhibited by DCA in vitro. Further examination of 13762 MAT rat mammary adenocarcinoma cells found that reversal of the glycolytic phenotype by DCA correlated with the inhibition of proliferation without any increase in cell death. This was despite a small but significant increase in caspase 3/7 activity, which may sensitize cancer cells to other apoptotic triggers. In vivo, DCA caused a 58% reduction in the number of lung metastases observed macroscopically after injection of 13762 MAT cells into the tail vein of rats (P = 0.0001, n > or = 9 per group). These results demonstrate that DCA has anti-proliferative properties in addition to pro-apoptotic properties, and can be effective against highly metastatic disease in vivo, highlighting its potential for clinical use.

Heparanase and vascular endothelial growth factor expression is increased in hypoxia-induced retinal neovascularization

Publisher: Association for Research in Vision and Ophthalmology (ARVO)

Date: 03-10-2012

DOI: 10.1167/IOVS.11-9144

Abstract: Heparanase and VEGF are related closely to angiogenesis in cancer. The purpose of our study was to evaluate the expression and correlation of heparanase and VEGF in hypoxia-induced retinal neovascularization. C57BL/6 oxygen-induced retinopathy (OIR) mice and human retinal microvascular endothelial cells (HRECs) were treated with the hypoxia mimetic agent cobalt chloride (CoCl₂), and in the presence of the heparanase inhibitor phosphomannopentaose sulfate (Muparfostat, PI-88). Heparanase activity was assayed in HRECs, and the expression of heparanase, VEGF protein and mRNA were evaluated by immunofluorescence, ELISA, Western blot, and real-time PCR while retinal flat mounts were used to evaluate the area of neovascularization of mice retina. HREC heparanase activity was increased by treatment with CoCl₂, but was decreased by PI-88. Immunofluorescence showed that heparanase and VEGF staining was intense in hypoxia-treated HRECs and OIR mice retina, while VEGF staining was faint in the normoxia and PI-88-treated ones. Western blot and real-time PCR results indicated that the expression of heparanase and VEGF was increased under hypoxic conditions, and the increase of VEGF was inhibited by PI-88. Retinal flat mounts showed that the area of new vessels in retina of OIR mice was increased compared to the normoxic mice, and this effect was inhibited by PI-88. Heparanase is upregulated and associated with the VEGF expression in hypoxia-induced retinal diseases. Heparanase is involved in hypoxia-induced neovascularization through promoting VEGF expression and may be a new therapeutic target for hypoxia-induced neovascularization retinal diseases.

Heme oxygenase-1 deficiency alters erythroblastic island formation, steady-state erythropoiesis and red blood cell lifespan in mice

Publisher: Ferrata Storti Foundation (Haematologica)

Date: 14-02-2015

DOI: 10.3324/HAEMATOL.2014.116368

Enhanced antibody responses to antigens presented on autologous erythrocytes

Publisher: Elsevier BV

Date: 12-1977

DOI: 10.1016/0022-1759(77)90179-X

Abstract: On a dose basis, antigen coupled to autologous red blood cells is 1,000--10,000-fold more efficient at inducing an antibody response than the soluble form. More than one antigen can be coupled simultaneously to the same red blood cells. Under these circumstances, prior immunization with one antigen enhances the antibody response to the other antigen, provided both antigens are coupled to the same red cell. Thus, the technique of coupling antigens to red blood cells is a means of producing high-titred antisera without the use of adjuvant and also represents a useful procedure for preparing composite antigens for probing cell-cell interaction in the immune response.

Ability of Different Chemically Modified Heparins To Potentiate the Biological Activity of Heparin-Binding Growth Factor 1 : Lack of Correlation with Growth Factor Binding

Publisher: American Chemical Society (ACS)

Date: 21-07-1992

DOI: 10.1021/BI00143A020

Abstract: A range of chemically modified heparins was examined for their ability to bind heparin-binding growth factor 1 (HBGF-1 acidic fibroblast growth factor) and potentiate the in vitro mitogenic and neurotrophic activity of HBGF-1. It was found that carboxyl-reduced heparin bound HBGF-1 as effectively as the native heparin molecule. Totally desulfated heparin and N-desulfated heparin lack HBGF-1-binding capacity, and substitution of the exposed amino group with acetyl or acetoacetyl groups only partially restored binding capacity, indicating that N-sulfates only play a limited role in growth factor binding. However, the failure of totally desulfated, N-resulfated heparin to interact with HBGF-1 demonstrated that N-sulfates alone are insufficient and ester sulfates are absolutely essential for HBGF-1 binding. In contrast, the ability of the modified heparins to potentiate the mitogenic activity of HBGF-1 correlated only to a limited extent with their affinity for HBGF-1. Thus, the carboxyl-reduced molecule which displayed similar affinity for HBGF-1 as native heparin was consistently less potent in augmenting mitogenesis. Similarly, the N-acetylated and the N-acetoacetylated species, which had much lower affinity for HBGF-1 than the carboxyl-reduced molecule, conferred similar biological activity to HBGF-1 whereas N-desulfated heparin, which was unable to bind growth factor, potentiated the mitogenic activity of HBGF-1 for both 3T3 and HUVE cells. In contrast, the neurotrophic activity of HBGF-1 was potentiated by modified heparin species which failed to bind HBGF-1 and were without activity in the mitogenic assays.(ABSTRACT TRUNCATED AT 250 WORDS)

Expansion of the editorial board of Immunology and Cell Biology

Publisher: Wiley

Date: 03-2009

DOI: 10.1038/ICB.2009.7

Activation of tumour cell ECM degradation by thrombin-activated platelet membranes: potentially a P-selectin and GPIIb/IIIa-dependent process

Publisher: Springer Science and Business Media LLC

Date: 16-05-2015

DOI: 10.1007/S10585-015-9722-5

Abstract: The promotion of tumour metastasis by platelets may occur through several mechanisms including the induction of a more metastatic phenotype in tumour cells and assisted extravasation of circulating tumour cells. Whilst the mechanisms underlying platelet-assisted extravasation have been extensively studied, much less attention has been paid to the mechanisms underlying platelet promotion of an aggressive phenotype within a tumour cell population. Herein, we demonstrate in vitro that MDA-MB-231 breast carcinoma cells incubated with washed thrombin-activated platelet membranes adopt a Matrigel-degrading phenotype in a dose- and contact time-dependent manner. The same phenotypic change was observed with three other human tumour cell lines of erse anatomical origin. Moreover, tumour cell lines that had been cultured with washed thrombin-activated platelet membranes had a greater metastatic capacity when injected into mice. This in vivo effect was reliant upon a co-incubation period of >2 h implying a mechanism involving more than platelet membrane binding that occurred within 5 min. Upon further investigation it was found that simultaneous blocking of the platelet-membrane proteins P-selectin and GPIIb/IIIa prevented interactions between platelet membranes and MDA-MB-231 cells but also significantly reduced the ability of tumour cells to degrade Matrigel. These results confirm that platelets induce a more aggressive phenotype in tumour cells but also identify the platelet proteins involved in this effect. P-selectin and GPIIb/IIIa also play a role in assisting tumour cell extravasation and, thus, are ideal targets for the therapeutic intervention of both stages of platelet-assisted metastasis.

Melanoma protective antitumor immunity activated by catalytic DNA

Publisher: Springer Science and Business Media LLC

Date: 29-05-2018

DOI: 10.1038/S41388-018-0306-0

Abstract: Melanoma incidence is increasing worldwide, and although drugs such as BRAF/MEK small-molecule inhibitors and immune checkpoint antibodies improve patient outcomes, most patients ultimately fail these therapies and alternative treatment strategies are urgently needed. DNAzymes have recently undergone clinical trials with signs of efficacy and no serious adverse events attributable to the DNAzyme. Here we investigated c-Jun expression in human primary and metastatic melanoma. We also explored the role of T cell immunity in DNAzyme inhibition of primary melanoma growth and the prevention of growth in non-treated tumors after the cessation of treatment in a mouse model. c-Jun was expressed in 80% of melanoma cells in human primary melanomas (n = 17) and in 83% of metastatic melanoma cells (n = 38). In contrast, c-Jun was expressed in only 11% of melanocytes in benign nevi (n = 24). Dz13, a DNAzyme targeting c-Jun/AP-1, suppressed both Dz13-injected and untreated B16F10 melanoma growth in the same mice, an abscopal effect relieved in each case by administration of anti-CD4/anti-CD8 antibodies. Dz13 increased levels of cleaved caspase-3 within the tumors. New, untreated melanomas grew poorly in mice previously treated with Dz13. Administration of anti-CD4/anti-CD8 antibodies ablated this inhibitory effect and the tumors grew rapidly. Dz13 inhibited c-Jun expression, reduced intratumoral vascularity (vascular lumina area defined by CD31 staining), and increased CD4

Identification of active-site residues of the pro-metastatic endoglycosidase heparanase

Publisher: American Chemical Society (ACS)

Date: 29-11-2000

DOI: 10.1021/BI002080P

Abstract: Heparanase is a beta-D-endoglucuronidase that cleaves heparan sulfate (HS) and has been implicated in many important physiological and pathological processes, including tumor cell metastasis, angiogenesis, and leukocyte migration. We report herein the identification of active-site residues of human heparanase. Using PSI-BLAST and PHI-BLAST searches of sequence databases, similarities were identified between heparanase and members of several of the glycosyl hydrolase families (10, 39, and 51) from glycosyl hydrolase clan A (GH-A), including strong local identities to regions containing the critical active-site catalytic proton donor and nucleophile residues that are conserved in this clan of enzymes. Furthermore, secondary structure predictions suggested that heparanase is likely to contain an (alpha/beta)(8) TIM-barrel fold, which is common to the GH-A families. On the basis of sequence alignments with a number of glycosyl hydrolases from GH-A, Glu(225) and Glu(343) of human heparanase were identified as the likely proton donor and nucleophile residues, respectively. The substitution of these residues with alanine and the subsequent expression of the mutant heparanases in COS-7 cells demonstrated that the HS-degrading capacity of both was abolished. In contrast, the alanine substitution of two other glutamic acid residues (Glu(378) and Glu(396)), both predicted to be outside the active site, did not affect heparanase activity. These data suggest that heparanase is a member of the clan A glycosyl hydrolases and has a common catalytic mechanism that involves two conserved acidic residues, a putative proton donor at Glu(225) and a nucleophile at Glu(343).

Ia antigenic specificities are oligosaccharide in nature: hapten-inhibition studies.

Publisher: Rockefeller University Press

Date: 04-1977

DOI: 10.1084/JEM.145.4.1039

Abstract: We have previously reported that the Ia specificities, coded for by the I region within the H-2 complex, appear to consist predominantly of carbohydrate. This conclusion was reached by examining low molecular weight Ia-bearing oligosacharides isolated from mouse serum. We now report hapten-inhibition studies which indicate that the binding of both allogeneic and xenogeneic anti-Ia antibodies to the Ia glycoproteins found predominantly on B lymphocytes can be specifically inhibited by certain free sugars. Both inhibition assays revealed that the specificity for the following Ia antigens resides predominantly in the following sugars: (a) Ia.1: N-acetyl-D-mannosamine or related sugars (b) Ia.3: alpha-D-galactose and related sugars (c) Ia.7: L-fucose and (d) Ia.15: N-acetyl-D-glucosamine. It seems likely that these sugars are found at the terminal nonreducing ends of the carbohydrate portion of the Ia-bearing glycoproteins present in the lymphocyte membrane. In contrast, several public and private H-2 antigenic specificities did not appear to be sugar defined. These studies imply that at least some of the Ia genes from both the I-A and I-C subregions of the I region code for glycosyl transferases which modify oligosaccharide structure and impart specificity to the Ia antigens by alteration of their terminal sugar residues.

Is host heparanase required for the rapid spread of heparan sulfate binding viruses?

Publisher: Elsevier BV

Date: 03-2019

DOI: 10.1016/J.VIROL.2019.01.001

Abstract: Vaccinia virus (VACV), like many other viruses, binds to cell surface heparan sulfate (HS) prior to infecting cells. Since HS is ubiquitously expressed extracellularly, it seemed likely that VACV-HS interaction may impede virus spread, with host heparanase, the only known mammalian endoglycosidase that can degrade HS, potentially overcoming this problem. In support of this hypothesis, we found that, compared to wild type, mice deficient in heparanase showed a 1-3 days delay in the spread of VACV to distant organs, such as ovaries, following intranasal inoculation, or to ovaries and spleen following intramuscular inoculation. These delays in spread occurred despite heparanase deficiency having no effect on VACV replication at inoculation sites. Subsequent in vitro studies revealed that heparanase treatment released VACV from HS expressing, but not HS deficient, infected cell monolayers. Collectively these data suggest that VACV relies on host heparanase to degrade HS in order to spread to distant sites.

Isolation and characterization of cell adhesion molecules from the marine sponge, Ophlitaspongia tenuis

Publisher: Elsevier BV

Date: 1991

DOI: 10.1016/0304-4165(91)90182-G

Abstract: Previous studies suggested that cell adhesion in the marine sponge, Ophlitaspongia tenuis, is mediated by a 35 kDa cell surface protein which interacts with an extracellular sulfated polysaccharide. This paper describes a simple and efficient procedure for isolating both putative cell adhesion molecules from detergent lysates of O. tenuis cells, the procedure being based on the fortuitous affinity of the sponge polysaccharide for heparin. The purified polysaccharide inhibits O. tenuis sponge cell aggregation, is highly sulfated and represents a glycosaminoglycan containing glucuronic acid. N-sulfated glucosamine and, possibly, glucose. The purified 35 kDa protein has a high affinity for the sponge polysaccharide and also, selectively interacts with dextran sulfate, a polysaccharide that has been shown previously to both bind to the sponge cell surface and inhibit aggregation of O. tenuis cells. Collectively, the data supports the hypothesis that the 35 kDa molecule is the major cell adhesion protein in O. tenuis. Preliminary data also suggests that the sponge contains an endogenous glycan hydrolase which can cleave the sponge polysaccharide.

Targeting dendritic cells with antigen-containing liposomes: Antitumour immunity

Publisher: Informa UK Limited

Date: 11-2004

DOI: 10.1517/14712598.4.11.1735

Abstract: Dendritic cells (DCs) are antigen-presenting cells that play an important role in the body's immune defence against cancer. Strategies using antigen-primed DCs as tumour vaccines show promise in patients, but the approach is cumbersome to use clinically. Soluble tumour antigens can be targeted to DCs in vivo, but this often induces antigenic tolerance rather than immunity. Liposomes are vesicular lipid structures with adjuvant-like properties. Importantly, liposomes can encapsulate antigen and immunomodulatory factors, thus serving as potent delivery vehicles. Different strategies are being explored to target liposomal antigens to DCs in vivo. One approach has employed single-chain antibody fragments to the DC surface molecules CD11c and DEC-205, attached to the vesicle surface by metal-chelating linkage, to target liposomal membranes containing antigen and either interferon-gamma or lipopolysaccharide to DCs. Such membranes induce dramatic antitumour responses and immunotherapeutic effects when used as a vaccine in the murine tumour model B16-OVA melanoma. Liposomal targeting of antigen and maturation signals directly to DCs in vivo, therefore, represents a much simpler strategy for cancer immunotherapy than antigen loading DCs ex vivo.

Inhibition of growth in vitro of plasmodium falciparum by complex polysaccharides

Publisher: Oxford University Press (OUP)

Date: 07-1988

DOI: 10.1016/0035-9203(88)90504-4

Abstract: Substitution of a metal center of phosphomolybdate, PMo(12)O(40) (3-) (PMo(12)), or its tungsten analogue with dirhodium(II) and subsequent stabilization of gold nanoparticles, AuNPs, with Rh(2)PMo(11) is demonstrated. The AuNP-Rh(2)PMo(11) mediates oxidations but adsorbs too weakly for direct modification of electrode materials. Stability in quiescent solution was achieved by modifying glassy carbon (GC) with 3-aminopropyltriethoxysilane (APTES) and then electrostatically assembling AuNP-Rh(2)PMo(11). At GC|APTES|AuNP-Rh(2)PMo(11), cyclic voltammetry showed the expected set of three reversible peak-pairs for PMo(11) in the range -0.2 to 0.6 vs (Ag/AgCl)/V and the reversible Rh(II,III) couple at 1.0 vs (Ag/AgCl)/V. The presence of AuNPs increased the current for the reduction of bromate by a factor of 2.5 relative to that at GC|Rh(2)PMo(11), and the electrocatalytic oxidation of methionine displayed characteristics of synergism between the AuNP and Rh(II). To stabilize AuNP-Rh(2)PMo(11) on a surface in a flow system, GC was modified by electrochemically assisted deposition of a sol-gel with templated 10-nm pores prior to immobilizing the catalyst in the pores. The resulting electrode permitted determination of bromate by flow-injection erometry with a detection limit of 4.0 × 10(-8) mol dm(-3).

Inhibition of secondary IgG responses by monosaccharides: evidence for I-region control.

Publisher: Wiley

Date: 04-1982

DOI: 10.1111/J.1744-313X.1982.TB00964.X

Abstract: Certain monosaccharides selectively inhibit secondary IgG responses in vitro. Genetic analyses described in this report revealed that the inhibitory sugars differed between mouse strains and these differences mapped to the I-J and I-C subregions of the murine MHC. These results imply that interaction between T and B lymphocytes can involve the recognition of I-region controlled carbohydrate structures.

Isolation and characterization of low molecular weight Ia-like antigens from normal human serum.

Publisher: Elsevier BV

Date: 06-1981

DOI: 10.1016/0161-5890(81)90129-2

Lipo-chitin oligosaccharides, plant symbiosis signalling molecules that modulate mammalian angiogenesis in vitro

Publisher: Public Library of Science (PLoS)

Date: 23-12-2014

DOI: 10.1371/JOURNAL.PONE.0112635

Estimating an optimal distribution of water entitlements

Publisher: American Geophysical Union (AGU)

Date: 12-1992

DOI: 10.1029/92WR01795

Extraction of the carbohydrate-defined class of Ia antigens from murine spleen cells and serum.

Publisher: Elsevier BV

Date: 08-1980

DOI: 10.1016/0161-5890(80)90101-7

Abstract: Although research has quantitatively evaluated the impacts of stigma on working women with disabilities (WWD), nuanced, qualitative accounts voiced by these women are rare. To address this literature gap, we conducted seven focus groups with forty-two WWD. We asked: "What are women's experiences of disability disclosure and accommodation in the workplace?" Findings reveal that WWD face intentional and unintentional structural discrimination and must weigh the pros and cons of disclosure and navigate devaluation threats in pursuing workplace accommodations. "Going the extra mile" emerged as a stigma management technique that was prevalent among women of higher social capital.

Astrocytic hypertrophy: An important pathological feature of chronic experimental autoimmune encephalitis in aged rats

Publisher: Elsevier BV

Date: 11-1993

DOI: 10.1016/0165-5728(93)90184-Z

Abstract: Experimental autoimmune encephalomyelitis (EAE) was induced in young (2-3 month old), middle-aged (12-13 month old) and geriatric (24-26 month old) Lewis (JC) rats by active immunisation with myelin basic protein (MBP) in complete Freund's adjuvant (CFA). It was found that aged Lewis (JC) rats developed a more chronic form of EAE than younger rats of the same strain, a phenomenon observed in both male and female rats despite males developing more severe disease than females at all ages. Middle-aged recipients also developed more severe disease than young recipients when EAE was induced by the adoptive transfer of lymphocytes from actively immunised young donors, suggesting that disease chronicity in middle-aged animals is a property of the central nervous system (CNS) milieu. Histological studies demonstrated that disease chronicity did not correlate with the number of inflammatory lesions in the CNS, young animals containing substantial numbers of CNS lesions following recovery and lesions being largely absent from middle-aged animals which still exhibited signs of disease. No significant differences were found in the degree of fibrin deposition or demyelination between young and middle-aged or symptomatic and asymptomatic animals. However, astrocytic hypertrophy was found to correlate with manifestation of disease in both young and middle-aged animals and in particular with disease chronicity in middle-aged animals. In parallel studies, no significant differences were found in the levels of the inflammatory mediators tumor necrosis factor (TNF)-alpha, prostaglandin E (PGE)2, reactive nitrogen intermediates (RNI) and corticosterone in young and middle-aged animals. However, markedly elevated corticosterone levels were found in both young and middle-aged animals with the development of clinical signs which returned to baseline levels with the resolution of clinical signs. Elevated levels of RNI were evident in animals immediately prior to and during the early stages of symptomatic EAE. Although these results suggest that nitric oxide may play a role in the pathogenesis of disease, whereas corticosterone may play a role in the immunoregulation of the disease, these factors cannot explain differences in disease chronicity evident in middle-aged animals.(ABSTRACT TRUNCATED AT 400 WORDS)

Automated fluorometric assay for T cell cytotoxicity

Publisher: Elsevier BV

Date: 08-1988

DOI: 10.1016/0022-1759(88)90042-7

Abstract: A fluorometric assay avoiding the use of radioactivity has been developed for detecting cytotoxic T lymphocytes (Tc cells). The method involves labelling targets with Hoechst dye no. 33342 (H33342) which becomes brightly fluorescent on binding to DNA. Lysis of target cells by Tc cells is quantified by measuring the release of fluorescent H33342 into the supernatant of culture wells. The fluorescence is measured using an automated Microfluor reader which allows results to be obtained rapidly. The assay has been used to detect alloreactive Tc cells and H-2 restricted Tc cells against influenza virus in a short-term 6 h assay using P815 and L929 as targets with comparable results to those obtained with 51Cr labelling. In contrast, lymphocyte blasts were found to be less sensitive in 6 h fluorometric assays when compared with the 51Cr assay. In long-term overnight assays (possible because of the low spontaneous release of H33342 from targets) lymphocyte blasts gave high specific lysis and some anti-self reactivity. The cause of the anti-self reactivity may reflect fundamental differences between the H33342 and 51Cr release assays.

New and improved methods for measuring lymphocyte proliferation in vitro and in vivo using CFSE-like fluorescent dyes

Publisher: Elsevier BV

Date: 05-2012

DOI: 10.1016/J.JIM.2012.02.012

Abstract: The use of carboxyfluorescein diacetate succinimidyl ester (CFSE) to measure lymphocyte proliferation by flow cytometry has become one of the most widely utilised assays for assessing lymphocyte responses. The properties of CFSE make it ideal for such a task, covalently labelling cells with a long-lived fluorescence of high intensity and low variance with minimal cell toxicity. No dye in the last 20 years has been capable of replicating CFSE in these respects. However, currently CFSE is limited to following a maximum of 7 cell isions and is not compatible for use with ubiquitously available fluorescein conjugates or other fluorescent molecules with spectral properties similar to fluorescein, such as EGFP. Here we characterise two new fluorescent dyes for measuring lymphocyte proliferation, Cell Trace Violet (CTV) and Cell Proliferation Dye eFluor 670 (CPD), which have different excitation and emission spectra to CFSE and, consequently, are compatible with fluorescein conjugates. We found that while both CTV and CPD can label cells to a high fluorescence intensity, which is long-lived and has low variability and low toxicity and makes them ideal for long-term tracking of non- iding lymphocytes in vivo, CTV offers possibly the best available alternative to CFSE in the analysis of cell isions. We also describe how intercellular dye transfer and cell autofluorescence can affect ision resolution with the three different dyes and describe labelling conditions for the three dyes that produce ultra-bright lymphocytes for in vivo tracking studies and allow up to 11 cell isions to be detected when using CFSE and CTV as the fluorescent dyes.

Heparan sulfate regulates IL-21 bioavailability and signal strength that control germinal center B cell selection and differentiation

Publisher: American Association for the Advancement of Science (AAAS)

Date: 24-02-2023

DOI: 10.1126/SCIIMMUNOL.ADD1728

Abstract: In antibody responses, mutated germinal center B (B GC ) cells are positively selected for reentry or differentiation. As the products from GCs, memory B cells and antibody-secreting cells (ASCs) support high-affinity and long-lasting immunity. Positive selection of B GC cells is controlled by signals received through the B cell receptor (BCR) and follicular helper T (T FH ) cell–derived signals, in particular costimulation through CD40. Here, we demonstrate that the T FH cell effector cytokine interleukin-21 (IL-21) joins BCR and CD40 in supporting B GC selection and reveal that strong IL-21 signaling prioritizes ASC differentiation in vivo. B GC cells, compared with non-B GC cells, show significantly reduced IL-21 binding and attenuated signaling, which is mediated by low cellular heparan sulfate (HS) sulfation. Mechanistically, N-deacetylase and N-sulfotransferase 1 (Ndst1)–mediated N-sulfation of HS in B cells promotes IL-21 binding and signal strength. Ndst1 is down-regulated in B GC cells and up-regulated in ASC precursors, suggesting selective desensitization to IL-21 in B GC cells. Thus, specialized biochemical regulation of IL-21 bioavailability and signal strength sets a balance between the stringency and efficiency of GC selection.

Glycosaminoglycan-induced activation of the β-secretase (BACE1) of Alzheimer’s disease

Publisher: Wiley

Date: 03-2010

DOI: 10.1111/J.1471-4159.2010.06571.X

Abstract: The beta-site APP cleaving enzyme (BACE1) is responsible for the first step in the production of the beta-amyloid protein of Alzheimer's disease. BACE1 is synthesized as a partially active zymogen (proBACE1). We previously showed that the glycosaminoglycan (GAG) heparin can increase the enzyme activity of proBACE1. In this study, the structural requirements and the mechanism for the GAG-induced activation were examined. The effect of heparin on proBACE1 was influenced by the degree of sulfation and carboxylation of the GAG, as well as by the length of the sugar. Although low molecular weight heparin fragments did not strongly stimulate proBACE1, they inhibited heparin-induced activation of the enzyme. The structure of the zymogen was modeled using the known X-ray structures of the BACE1 catalytic domain and the homologous prodomain of porcine pepsinogen. The modeled structure suggested that a heparin-binding domain may reside close to the prodomain, and that movement of a loop region between residues 46-65, lying adjacent to the prodomain, may be needed to accommodate heparin binding. The presence of the loop domain adjacent to the active site may account for the lower activity of the zymogen relative to the mature enzyme. Movement of the loop region upon heparin binding could expose the active site region to allow for increased substrate binding. The results suggest a model in which conformational changes close to the prodomain may be involved in the mechanism of heparin-induced activation of proBACE1.

Acute T-Cell-Driven Inflammation Requires the Endoglycosidase Heparanase-1 from Multiple Cell Types

Publisher: MDPI AG

Date: 21-04-2022

DOI: 10.3390/IJMS23094625

Abstract: It has been accepted for decades that T lymphocytes and metastasising tumour cells traverse basement membranes (BM) by deploying a battery of degradative enzymes, particularly proteases. However, since many redundant proteases can solubilise BM it has been difficult to prove that proteases aid cell migration, particularly in vivo. Recent studies also suggest that other mechanisms allow BM passage of cells. To resolve this issue we exploited heparanase-1 (HPSE-1), the only endoglycosidase in mammals that digests heparan sulfate (HS), a major constituent of BM. Initially we examined the effect of HPSE-1 deficiency on a well-characterised adoptive transfer model of T-cell-mediated inflammation. We found that total elimination of HPSE-1 from this system resulted in a drastic reduction in tissue injury and loss of target HS. Subsequent studies showed that the source of HPSE-1 in the transferred T cells was predominantly activated CD4+ T cells. Based on bone marrow chimeras, two cellular sources of HPSE-1 were identified in T cell recipients, one being haematopoiesis dependent and the other radiation resistant. Collectively our findings unequivocally demonstrate that an acute T-cell-initiated inflammatory response is HPSE-1 dependent and is reliant on HPSE-1 from at least three different cell types.

Lymphocytes express a diverse array of specific receptors for sulfated polysaccharides

Publisher: Elsevier BV

Date: 03-1985

DOI: 10.1016/0008-8749(85)90044-9

Abstract: Lymphocyte receptors for sulfated polysaccharides were detected in two ways, namely, by the ability of lymphocytes to form rosettes with sheep red blood cells (SRBC) coupled with one of fourteen different sulfated polysaccharides, and by the ability of cholate extracts of lymphocytes to hemagglutinate the same sulfated polysaccharide-coupled SRBC. It was found that murine lymphocytes lacked receptors for a number of glycosaminoglycans, such as hyaluronic acid, chondroitin-4-sulfate, chondroitin-6-sulfate, and dermatan sulfate, but reacted strongly with heparin, arteparon, and a number of sulfated polysaccharides of plant and bacterial origin. In each case receptor activity was demonstrated by rosetting and by the ability of lymphocyte lysates to strongly agglutinate sulfated polysaccharide-coupled SRBC. The receptors exhibited a high degree of ersity as evidenced by (a) only subpopulations of lymphocytes, particularly splenic B cells, expressing receptors for some of the sulfated polysaccharides and (b) hemagglutination-inhibition analyses revealing numerous subsets of receptors with different binding specificities. Receptor ersity was further highlighted by a 48% difference in the hemagglutination-inhibiton results between thymus and spleen. It is proposed that these receptors are involved in cell-cell communication and lymphocyte homing and recirculation. The likely target structures for the receptors in vivo are the heparan sulfates, a ubiquitous and structurally erse family of sulfated glycosaminoglycans.

Regulation of the immune response II. Repressor T cells in cyclophosphamide-induced tolerant mice

Publisher: Wiley

Date: 03-1977

DOI: 10.1002/EJI.1830070313

Abstract: Between August 1981 and February 1987, 67 orthotopic heart transplants and three heart-lung transplants were performed in 69 patients at the University of Munich Hospital. The immunosuppressive regimen consisted of cyclosporine A, azathioprine, and prednisone. The diagnosis of acute rejection was based on cytoimmunologic monitoring, frequency analysis of fast Fourier transformed surface electrocardiograms (FFT-ECGs), and two-dimensional echocardiography. The results of these diagnostic methods were compared to the findings provided by endomyocardial biopsies, which were performed simultaneously with the noninvasive studies. Seventy patients underwent cytoimmunologic monitoring. In 88% of all rejection episodes, this technique revealed activated lymphocytes and lymphoblasts in the mononuclear concentrate of the peripheral blood s les the presence of such cells is known to be an extremely early sign of acute rejection. Twenty-six patients were monitored by means of FFT-ECG. In 20 of the 21 cases of rejection, this method disclosed significant changes in the frequency spectrum of the QRS complex in the 70- to 110-Hz range in 12 cases, these changes were the earliest sign of acute rejection. Therefore, FFT-ECG had a sensitivity of 95%. All of the QRS changes were reversible with rejection therapy. Forty-five patients were subjected to two-dimensional echocardiography. In 31 of the 35 cases of rejection, the echocardiogram showed a significant increase in the left ventricular wall thickness and a decrease in the left ventricular cross-sectional area during mild rejection. Moderate or severe rejection was characterized by an increase in the diastolic area, as well as a decrease in the systolic area change and in the diastolic maximum velocity of area change. Thus, two-dimensional echocardiography had a sensitivity of 89%. In the recent cases, the diagnosis of rejection was based on noninvasive methods alone. After rejection therapy had been instituted, endomyocardial biopsies were performed to assess the effectiveness of the treatment. With noninvasive rejection monitoring, the number of endomyocardial biopsies performed during the first three postoperative months was only 2.8 per patient in comparison with invasive rejection monitoring, noninvasive follow-up was associated with a 75% reduction in the need for biopsy.

DNAzyme targeting c-jun suppresses skin cancer growth

Publisher: American Association for the Advancement of Science (AAAS)

Date: 20-06-2012

DOI: 10.1126/SCITRANSLMED.3003960

Abstract: Catalytic DNA molecules that target the transcription factor c- jun inhibit skin cancer growth in mice.

Targeting RNA Polymerase I Transcription Activity in Osteosarcoma: Pre-Clinical Molecular and Animal Treatment Studies

Publisher: MDPI AG

Date: 09-04-2023

DOI: 10.3390/BIOMEDICINES11041133

Abstract: The survival rate of patients with osteosarcoma (OS) has not improved over the last 30 years. Mutations in the genes TP53, RB1 and c-Myc frequently occur in OS and enhance RNA Polymerase I (Pol I) activity, thus supporting uncontrolled cancer cell proliferation. We therefore hypothesised that Pol I inhibition may be an effective therapeutic strategy for this aggressive cancer. The Pol I inhibitor CX-5461 has demonstrated therapeutic efficacy in different cancers in pre-clinical and phase I clinical trials thus, the effects were determined on ten human OS cell lines. Following characterisation using genome profiling and Western blotting, RNA Pol I activity, cell proliferation and cell cycle progression were evaluated in vitro, and the growth of TP53 wild-type and mutant tumours was measured in a murine allograft model and in two human xenograft OS models. CX-5461 treatment resulted in reduced ribosomal DNA (rDNA) transcription and Growth 2 (G2)-phase cell cycle arrest in all OS cell lines. Additionally, tumour growth in all allograft and xenograft OS models was effectively suppressed without apparent toxicity. Our study demonstrates the efficacy of Pol I inhibition against OS with varying genetic alterations. This study provides pre-clinical evidence to support this novel therapeutic approach in OS.

Determination of lymphocyte division by flow cytometry

Publisher: Elsevier BV

Date: 05-1994

DOI: 10.1016/0022-1759(94)90236-4

Abstract: Techniques currently available for determining cell ision are able to show one or, at best, a limited number of cell isions. Other methods exist which can quantify overall ision, but tell nothing about the ision history of in idual cells. Here we present a new technique in which an intracellular fluorescent label is ided equally between daughter cells upon cell ision. The technique is applicable to in vitro cell ision, as well as in vivo ision of adoptively transferred cells, and can resolve multiple successive generations using flow cytometry. The label is fluorescein derived, allowing monoclonal antibodies conjugated to phycoerythrin or other compatible fluorochromes to be used to immunophenotype the iding cells.

Eotaxin Selectively Binds Heparin

Publisher: Elsevier BV

Date: 05-2007

DOI: 10.1074/JBC.M608046200

Glycosyltransferases and T-cell recognition.

Publisher: Elsevier BV

Date: 06-1981

DOI: 10.1016/0167-5699(81)90040-2

Abstract: Recent studies suggest that the AIIIC locus controls families of carbohydrate, as well as protein, histocompatibility antigens. In this article Christopher Parish and his colleagues discuss the molecular and genetic implications of these findings and present the hypothesis that the glycosyltransferase enzymes which construct the carbohydrate histocompatibility antigens may be involved in T-cell recognition.

The Role of Th2-Mediated Anti-Tumor Immunity in Tumor Surveillance and Clearance

Publisher: Humana Press

Date: 31-10-2009

DOI: 10.1007/978-1-60761-451-7_11

Autorosette inhibition factor: isolation and properties of the human plasma protein.

Publisher: Wiley

Date: 10-1981

DOI: 10.1111/J.1432-1033.1981.TB05655.X

Binding sites for glycosaminoglycans on developing sympathetic neurones

Publisher: Wiley

Date: 1986

DOI: 10.1002/JNR.490150407

Abstract: Recent studies have suggested that cell to cell communication in the immune system is mediated by cell surface receptors for glycosaminoglycans (GAGs) [Parish et al, 1984]. The intention of this study was to see whether similar recognition molecules for GAGs are present on sympathetic neurones. A mechanical dissociation technique was used to isolate neurones from superior cervical ganglia (SCG) of rats aged between gestational day 19 and postnatal day 21. Receptors for GAGs on sympathetic neurones were detected by the ability of neurones to form rosettes with sheep red blood cells coupled with one of 12 different GAGs. It was found that SCG cells bind to all the GAGs tested. In addition, a range of developmental binding patterns for the various GAGs was found.

Low molecular weight Ia antigens in normal mouse serum

Publisher: Springer Science and Business Media LLC

Date: 12-1976

DOI: 10.1007/BF01576944

Low molecular weight Ia antigens in normal mouse serum

Publisher: Springer Science and Business Media LLC

Date: 12-1976

DOI: 10.1007/BF01576945

Synthesis, Structural Characterisation, and Preliminary Evaluation of Non-Indolin-2-one-based Angiogenesis Inhibitors Related to Sunitinib (Sutent®)

Publisher: CSIRO Publishing

Date: 2013

DOI: 10.1071/CH13219

Abstract: The indolin-2-one fused-ring system and the 2,4-dimethylpyrrole unit represent key structural motifs in the anticancer drug sunitinib (Sutent®) and predecessor angiogenesis inhibitors that have undergone anticancer clinical trials (e.g. semaxanib, SU5416). In pursuit of novel anti-angiogenic scaffolds, we were interested in identifying whether the indolin-2-one group in these structures could be modified without losing activity. This paper describes novel condensation chemistry used to prepare a test series of (E)- and (Z)-alkenes related to SU5416 that retain the 2,4-dimethylpyrrole unit while incorporating ring-opened indolin-2-ones. Unique structural characteristics were identified in the compounds, such as intramolecular hydrogen bonds in the (Z)-alkenes, and several ex les were shown to possess significant anti-angiogenic activity in a rat aorta in vitro model of angiogenesis. The work demonstrates that the indolin-2-one moiety is not an absolute requirement for angiogenesis inhibition in the sunitinib/SU5416 class.

New insights into intracellular locations and functions of heme oxygenase-1

Publisher: Mary Ann Liebert Inc

Date: 10-04-2014

DOI: 10.1089/ARS.2013.5675

Beware of NK cells in pre-clinical metastasis models

Publisher: Springer Science and Business Media LLC

Date: 19-04-2013

DOI: 10.1007/S10585-013-9582-9

Functional significance of the complement receptors on B lymphocytes

Publisher: Elsevier BV

Date: 12-1975

DOI: 10.1016/0008-8749(75)90106-9

Detection of low-affinity adhesion ligands by linking recombinant cell adhesion molecules in uniform orientation to a fluorescently labelled dextran molecule by means of hexahistidine tagging: The case of multimeric CD40

Publisher: Elsevier BV

Date: 02-1998

DOI: 10.1016/S0022-1759(98)00030-1

Abstract: Cell-cell interactions involve highly polyvalent associations between receptors on adjacent cells. In order to mimic this process, we have prepared a highly polyvalent form of CD40 attached to a dextran backbone. This was accomplished by engineering a hexahistidine tag on the C-terminus of the CD40 and binding, in a uniform orientation, up to 100 molecules of hexahistidine CD40 by metal chelation to a single fluorescently tagged dextran molecule. The advantage of this 'multimeric' CD40 is that it would be expected to bind to any counterstructure with a significantly higher avidity compared to monomeric CD40. The multimeric CD40 bound with high affinity to stably transfected mouse fibroblasts expressing CD40L. The multimeric ligand also bound to the activated T cell clone, D10, but did not bind to resting cells, showing that it bound to the physiological ligand. Using this system, we found no evidence to support the claim [Heath et al., 1993. Cell. Immunol. 152, 468.] that the A20 cells have a counterstructure for CD40, and propose that the high binding of CD40 observed in this study may have been due to an exposed hexahistidine tag on the molecule. This multimeric technology has considerable potential for detecting low-affinity interactions between cell adhesion receptors and ligands. The uniform orientation of the molecules on the dextran is an advantage over previous systems and permits the preparation of heterogeneous, multimeric ligands which more closely mimic the conditions at the cell surface.

All soft contact lenses are not created equal

Publisher: Elsevier BV

Date: 04-2022

DOI: 10.1016/J.CLAE.2021.101515

Sulfated polyanions prevent HIV infection of lymphocytes by disruption of the CD4-gp120 interaction, but do not inhibit monocyte infection

Publisher: Oxford University Press (OUP)

Date: 09-1994

DOI: 10.1002/JLB.56.3.266

Abstract: Sulfated polyanions (SPs) bind variably to lymphocyte-expressed CD4 and inhibit binding of monoclonal antibodies to the first two domains of CD4. To further define this interaction, soluble recombinant CD4 (sCD4 four extracellular domains), its truncated amino-terminal two-domain derivative, and three linear peptide analogues spanning residues 6–60 (6–24, 20–40, 41–60) in the first domain were investigated for SP binding. Dextran sulfate (DXS) (500 kDa), polyvinyl sulfate, fucoidan, and carrageenan-kappa, each immobilized on carboxymethyl cellulose fibers, bound strongly to both the two-domain and four-domain recombinant CD4 molecules (similar to that observed with native CD4), whereas dextran sulfate (5 kDa), chondroitin 6-sulfate, and pentosan sulfate bound relatively poorly. No peptide binding to SPs was observed. Recombinant gp120 bound poorly (& %) to all of the immobilized polyanions, except pentosan sulfate (17%), for which some binding was noted. Binding of radiolabeled V3 loop peptide to SPs was slightly greater, with 20-30% binding to polyvinyl sulfate, dextran sulfate (500 kDa), and pentosan sulfate. Competitive binding studies demonstrated the predominance of sCD4 rather than rgp120 binding to SPs and supported previous data demonstrating a binding site for DXS (500 kDa) on the first domain of CD4 adjacent to the gp120 binding site and recognized by OKT4C and E monoclonal antibodies. Hence disruption of the CD4-gp120 interaction is probably responsible for most of the observed antiviral activity of SPs toward HIV infection of lymphocytes. However, HIV infection and gp120 binding to monocytes was unaffected by SPs, probably because SPs were unable to block the CD4-gp 120 interaction in monocytes. J. Leukoc. Biol. 56: 266–272 1994.

A one-step procedure for separating mouse T and B lymphocytes

Publisher: Wiley

Date: 12-1974

DOI: 10.1002/EJI.1830041208

Developing neolignans as proangiogenic agents: Stereoselective total syntheses and preliminary biological evaluations of the four guaiacylglycerol 8-O-4'-coniferyl ethers

Publisher: American Chemical Society (ACS)

Date: 30-10-2017

DOI: 10.1021/ACSOMEGA.7B01459

C-Glycoside formation via Lewis acid promoted reaction of O-glycosylimidates with pyrroles

Publisher: Royal Society of Chemistry (RSC)

Date: 09-07-2002

DOI: 10.1039/B205690A

Convergent synthesis and preliminary biological evaluations of the stilbenolignan (±)-aiphanol and various congeners

Publisher: Royal Society of Chemistry (RSC)

Date: 2003

DOI: 10.1039/B305106D

Abstract: Treatment of an equimolar mixture of stilbene 7 and cinnamyl alcohol 8 with silver carbonate in acetone-benzene afforded a ca. 2:1:2:1 mixture of the stilbenolignan (+/-)-aiphanol (1) and congeners 2-4 each of which show significant anti-angiogenic and COX-2 inhibitory properties.

Bystander B cells rapidly acquire antigen receptors from activated B cells by membrane transfer

Publisher: Proceedings of the National Academy of Sciences

Date: 18-03-2008

DOI: 10.1073/PNAS.0800259105

Abstract: The B cell antigen receptor (BCR) efficiently facilitates the capture and processing of a specific antigen for presentation on MHC class II molecules to antigen-specific CD4 + T cells ( 1 ). Despite this, the majority of B cells are thought to play only a limited role in CD4 + T cell activation because BCRs are clonotypically expressed. Here, we show, however, that activated B cells can, both in vitro and in vivo , rapidly donate their BCR to bystander B cells, a process that is mediated by direct membrane transfer between adjacent B cells and is lified by the interaction of the BCR with a specific antigen. This results in a dramatic expansion in the number of antigen-binding B cells in vivo , with the transferred BCR endowing recipient B cells with the ability to present a specific antigen to antigen-specific CD4 + T cells.

Neutralizing the pathological effects of extracellular histones with small polyanions

Publisher: Springer Science and Business Media LLC

Date: 16-12-2029

DOI: 10.1038/S41467-020-20231-Y

Abstract: Extracellular histones in neutrophil extracellular traps (NETs) or in chromatin from injured tissues are highly pathological, particularly when liberated by DNases. We report the development of small polyanions (SPAs) (~0.9–1.4 kDa) that interact electrostatically with histones, neutralizing their pathological effects. In vitro, SPAs inhibited the cytotoxic, platelet-activating and erythrocyte-damaging effects of histones, mechanistic studies revealing that SPAs block disruption of lipid-bilayers by histones. In vivo, SPAs significantly inhibited sepsis, deep-vein thrombosis, and cardiac and tissue-flap models of ischemia-reperfusion injury (IRI), but appeared to differ in their capacity to neutralize NET-bound versus free histones. Analysis of sera from sepsis and cardiac IRI patients supported these differential findings. Further investigations revealed this effect was likely due to the ability of certain SPAs to displace histones from NETs, thus destabilising the structure. Finally, based on our work, a non-toxic SPA that inhibits both NET-bound and free histone mediated pathologies was identified for clinical development.

The influence of platelet membranes on tumour cell behaviour.

Publisher: Springer Science and Business Media LLC

Date: 06-2017

DOI: 10.1007/S10555-017-9671-3

Abstract: The significant role of platelets in the protection of tumour cells from immune attack and shear forces and the promotion of tumour cell extravasation from the bloodstream in the process of haematogenous metastasis have been extensively studied. The role of platelets, and in particular platelet membranes, in the promotion of a more metastatic phenotype in tumour cells is a more recent and, therefore, less well-recognised area of research. This review article summarises studies that have focused on the impact of tumour cell interactions with platelets and platelet membranes on tumour cell behaviour in vitro and in vivo. Furthermore, the gene expression changes that occur within tumour cells following contact with platelet membranes are also extensively reviewed. Overall, the interaction of platelet membranes with tumour cells results in a more invasive phenotype and the promotion of epithelial to mesenchymal transition with our own genetic studies revealing that matrix metalloproteinase-1, plasminogen activator inhibitor-1 and interleukin-8 are globally upregulated in a range of tumour cell lines.

The uptake of soluble and nanoparticulate imaging isotope in model liver tumours after intra-venous and intra-arterial administration

Publisher: Elsevier BV

Date: 2015

DOI: 10.1016/J.BIOMATERIALS.2014.11.001

Abstract: Delivery of chemotherapeutic drugs to tumours by reformulation as nanoparticles has often been proposed as a means of facilitating increased selective uptake, exploiting the increased permeability of the tumour vasculature. However realisation of this improvement in drug delivery in cancer patients has met with limited success. We have compared tumour uptake of soluble Tc99m-pertechnetate and a colloid of nanoparticles with a Tc99m core, using both intra-venous and intra-arterial routes of administration in a rabbit liver VX2 tumour model. The radiolabelled nanoparticles were tested both in untreated and cationised form. The results from this tumour model in an internal organ show a marked advantage in intra-arterial administration over the intra-venous route, even for the soluble isotope. Tumour accumulation of nanoparticles from arterial administration was augmented by cationisation of the nanoparticle surface with histone proteins, which consistently facilitated selective accumulation within microvessels at the periphery of tumours.

Heparan sulfate and heparanase play key roles in mouse β cell survival and autoimmune diabetes

Publisher: American Society for Clinical Investigation

Date: 03-01-2012

DOI: 10.1172/JCI46177

Liposomal vaccines-targeting the delivery of antigen

Publisher: Elsevier BV

Date: 09-2006

DOI: 10.1016/J.YMETH.2006.05.027

Abstract: Vaccines that can prime the adaptive immune system for a quick and effective response against a pathogen or tumor cells, require the generation of antigen (Ag)-specific memory T and B cells. The unique ability of dendritic cells (DCs) to activate naïve T cells, implies a key role for DCs in this process. The generation of tumor-specific CD8(+) cytotoxic T cells (CTLs) is dependent on both T cell stimulation with Ag (peptide-MHC-complexes) and costimulation. Interestingly, tumor cells that lack expression of T cell costimulatory molecules become highly immunogenic when transfected to express such molecules on their surface. Adoptive immunotherapy with Ag-pulsed DCs also is a strategy showing promise as a treatment for cancer. The use of such cell-based vaccines, however, is cumbersome and expensive to use clinically, and/or may carry risks due to genetic manipulations. Liposomes are particulate vesicular lipid structures that can incorporate Ag, immunomodulatory factors and targeting molecules, and hence can serve as potent vaccines. Similarly, Ag-containing plasma membrane vesicles (PMV) derived from tumor cells can be modified to incorporate a T cell costimulatory molecule to provide both TCR stimulation, and costimulation. PMVs also can be modified to contain IFN-gamma and molecules for targeting DCs, permitting delivery of both Ag and a DC maturation signal for initiating an effective immune response. Our results show that use of such agents as vaccines can induce potent anti-tumor immune responses and immunotherapeutic effects in tumor models, and provide a strategy for the development of effective vaccines and immunotherapies for cancer and infectious diseases.

Regulation of the immune response by antibody. II. The effect of different classes of passive guinea pig antibody on humoral and cell-mediated immunity in rats

Publisher: Elsevier BV

Date: 12-1972

DOI: 10.1016/0008-8749(72)90101-3

Abstract: Nucleolar and spindle-associated protein 1 (NUSAP1) is an important mitotic regulator. In addition to its crucial function in mitosis, NUSAP1 has recently received attention due to the interesting roles in carcinogenesis. The aim of this study was to reveal functional mechanisms of NUSAP1 in oral squamous cell carcinoma (OSCC). mRNA and protein expression levels of NUSAP1 in 9 OSCC-derived cells were analyzed by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) and immunoblotting analyses. The correlation between the NUSAP1 expression profile and the clinicopathological factors was evaluated by immunohistochemistry (IHC) in clinical OSCC s les (n = 70). The NUSAP1 knockdown cells were established with short hairpin RNA (shRNA) in OSCC cells, and functional assays were performed using these cells. In addition to the evaluation of cellular proliferation and cell cycle, we also investigated the potential role of NUSAP1 in paclitaxel (PTX)-induced cellular responses. mRNA and protein expression of NUSAP1 were significantly up-regulated in OSCC-derived cells compared with human normal oral keratinocytes (P < 0.05). IHC revealed that NUSAP-1 expression is closely associated with primary advanced T stage (P<0.05). Suppression of NUSAP1 expression levels led to significant (P < 0.05) inhibition of cellular proliferation. Furthermore, apoptosis induced by PTX was enhanced in NUSAP1 knockdown OSCC cells. NUSAP1 may be a crucial biomarker for OSCC. Moreover, down-regulated NUSAP1 expression suppresses tumor proliferation and also enhances anti-tumor effect of PTX by activating apoptotic pathways. Thus, the present study strongly suggests that regulating NUSAP1 expression should contribute to the therapy for OSCC.

Immunology and Cell Biology joins Nature Publishing

Publisher: Wiley

Date: 2007

DOI: 10.1038/SJ.ICB.7100017

Regulation of Carcinogenesis by IL-5 and CCL11: A Potential Role for Eosinophils in Tumor Immune Surveillance

Publisher: The American Association of Immunologists

Date: 04-2007

DOI: 10.4049/JIMMUNOL.178.7.4222

Abstract: The role of the immune system in the surveillance of transformed cells has seen a resurgence of interest in the last 10 years, with a substantial body of data in mice and humans supporting a role for the immune system in host protection from tumor development and in shaping tumor immunogenicity. A number of earlier studies have demonstrated that eosinophils, when recruited into tumors, can very effectively eradicate transplantable tumors. In this study, we investigated whether eosinophils also play a role in tumor immune surveillance by determining the incidence of methylcholanthrene (MCA)-induced fibrosarcomas in IL-5 transgenic mice that have greatly enhanced levels of circulating eosinophils, CCL11 (eotaxin-1)-deficient mice that lack a key chemokine that recruits eosinophils into tissues, and the eosinophil-deficient mouse strains, IL-5/CCL11−/− and ΔdblGATA. It was found that MCA-induced tumor incidence and growth were significantly attenuated in IL-5 transgenic mice of both the BALB/c and C57BL/6 backgrounds. Histological examination revealed that the protective effect of IL-5 was associated with massively enhanced numbers of eosinophils within and surrounding tumors. Conversely, there was a higher tumor incidence in CCL11−/− BALB/c mice, which was associated with a reduced eosinophil influx into tumors. This correlation was confirmed in the eosinophil-deficient IL-5/CCL11−/− and ΔdblGATA mouse strains, where tumor incidence was greatly increased in the total absence of eosinophils. In addition, subsequent in vitro studies found that eosinophils could directly kill MCA-induced fibrosarcoma cells. Collectively, our data support a potential role for the eosinophil as an effector cell in tumor immune surveillance.

Evidence that the cellular ligand for the human NK cell activation receptor NKp30 is not a heparan sulfate glycosaminoglycan

Publisher: The American Association of Immunologists

Date: 07-2005

DOI: 10.4049/JIMMUNOL.175.1.207

Abstract: NKp30 (NCR3, CD337) is a natural cytotoxicity receptor, expressed on subsets of human peripheral blood NK cells, involved in NK cell killing of tumor cells and immature dendritic cells. The cellular ligand for NKp30 has remained elusive, although evidence that membrane-associated heparan sulfate (HS) proteoglycans are involved in the recognition of cellular targets by NKp30 was recently reported. The data presented in this report show conclusively that HS glycosaminoglycans (GAG) are not ligands for NKp30. We show that removing HS completely from the cell surface of human 293-EBNA cells with mammalian heparanase does not affect binding of rNKp30/human IgG1 Fc chimera complexes or binding of multimeric liposome-rNKp30 complexes. Removing HS from 293-EBNA cells, culture-generated DC, MM-170 malignant melanoma cells, or HeLa cells does not affect the NKp30-dependent killing of these cells by NK cells. We show further that the GAG-deficient hamster pgsA-745 cells that lack HS and the GAG-expressing parent CHO-K1 cells are both killed by NK cells, with killing of both cell lines inhibited to the same extent by anti-NKp30 mAb. From these results we conclude that HS GAG are not ligands for NKp30, leaving open the question as to the nature of the cellular ligand for this important NK cell activation receptor.

Unexpected new roles for heparanase in Type 1 diabetes and immune gene regulation

Publisher: Elsevier BV

Date: 06-2013

DOI: 10.1016/J.MATBIO.2013.02.007

Abstract: Heparanase (Hpse) is an endo-β-d-glucuronidase that degrades the glycosaminoglycan heparan sulfate (HS) in basement membranes (BMs) to facilitate leukocyte migration into tissues. Heparanase activity also releases HS-bound growth factors from the extracellular matrix (ECM), a function that aids wound healing and angiogenesis. In disease states, the degradation of HS in BMs by heparanase is well recognized as an invasive property of metastatic cancer cells. Recent studies by our group, however, have identified unexpected new roles for heparanase and HS. First, we discovered that in Type 1 diabetes (T1D) (i) HS in the pancreatic islet BM acts as a barrier to invading cells and (ii) high levels of HS within the insulin-producing islet beta cells themselves are critical for beta cell survival, protecting the cells from free radical-mediated damage. Furthermore, catalytically active heparanase produced by autoreactive T cells and other insulitis mononuclear cells was shown to degrade intra-islet HS, increasing the susceptibility of islet beta cells to free radical damage and death. This totally novel molecular explanation for the onset of T1D diabetes opens up new therapeutic approaches for preventing disease progression. Indeed, administration of the heparanase inhibitor, PI-88, dramatically reduced T1D incidence in diabetes-prone NOD mice, preserved islet beta cell HS and reduced islet inflammation. Second, in parallel studies it has been shown that heparanase and HS can be transported to the nucleus of cells where they impact directly or indirectly on gene transcription. Based on ChIP-on-chip studies heparanase was found to interact with the promoters and transcribed regions of several hundred genes and micro-RNAs in activated Jurkat T cells and up-regulate transcription, with many of the target genes/micro-RNAs being involved in T cell differentiation. At the molecular level, nuclear heparanase appears to regulate histone 3 lysine 4 (H3K4) methylation by influencing the recruitment of demethylases to transcriptionally active genes. These studies have unveiled new functions for heparanase produced by T lymphocytes, with the enzyme mediating unexpected intracellular effects on T cell differentiation and insulin-producing beta cell survival in T cell-dependent autoimmune T1D.

Alternatively Activated Macrophage Possess Antitumor Cytotoxicity That Is Induced by IL-4 and Mediated by Arginase-1

Publisher: Ovid Technologies (Wolters Kluwer Health)

Date: 06-2010

DOI: 10.1097/CJI.0B013E3181CD8746

Ia antigens in serum during different murine infections.

Publisher: American Society for Microbiology

Date: 11-1979

DOI: 10.1128/IAI.26.2.422-426.1979

Abstract: There exists in the mouse a family of I-region-controlled (Ia) antigens which carry carbohydrate-defined determinants. These antigens appear in serum as glycolipids and seem to be actively secreted by antigen-activated T-cells. This paper describes the ability of selected viral, bacterial, and protozoal infections of mice to markedly alter the serum levels of these Ia antigens. All the infectious agents examined induced substantial augmentation or suppression of serum Ia concentrations or both. Lymphocytic choriomeningitis (LCM) virus first enhanced and then suppressed serum Ia levels during the course of acute infection. Enhancement occurred during the time of ongoing virus replication and splenic lymphoproliferation while suppression coincided with the peak of the cytotoxic T-cell response and virus clearance. Listeria monocytogenes infection induced a substantial reduction in Ia levels at a time just after marked depletion of T-cells in the spleen. In contrast, Brucella abortus caused a significant increase in Ia levels 7 days postinfection, which correlates with the appearance of peak numbers of bacteria in tissues. Finally, Plasmodium yoelii, a nonlethal malarial parasite which stimulates prolonged T-cell proliferation, augmented serum Ia levels, whereas P. berghei, a lethal parasite which tends to inhibit. T-cell ision, suppressed Ia secretion. Possible interpretations of these different results are presented.

Optimal Time Selection for the Polarisation and Depolarisation Current Measurement for Power Transformer Insulation Diagnosis

Publisher: IEEE

Date: 06-2007

DOI: 10.1109/PES.2007.385514

Yin Yang-1 inhibits tumor cell growth and inhibits p21 ^WAF1/Cip1complex formation with cdk4 and cyclin D1

Publisher: Spandidos Publications

Date: 09-02-2012

DOI: 10.3892/IJO.2012.1362

Abstract: The GLI-Krüppel zinc finger factor yin yang-1 (YY1) is a complex protein that regulates a variety of processes including transcription, proliferation, development and differentiation. YY1 inhibits cell growth in a cell type-specific manner. The role played by YY1 in its control of tumor cell growth is unclear and controversial. We show here that YY1 can suppress the growth of different tumor cell types in vitro, including human breast carcinoma cells and glioblastoma cells. YY1 also blocked the growth of 13762 MAT mammary adenocarcinoma isografts in rats. YY1 inhibited 13762 MAT tumor growth by approximately 80% compared with the GFP alone group 21 days after injection. YY1 inhibited proliferating cell nuclear antigen (PCNA) expression and pRbSer249/Thr252 phosphorylation without influencing tumor microvascular density. Moreover, YY1 inhibited p21WAF1/Cip1 complex formation with cdk4 and cyclin D1. These findings demonstrate that YY1 can negatively regulate the growth of multiple malignant cell types.

Histidine-rich glycoprotein prevents the formation of insoluble immune complexes by rheumatoid factor

Publisher: Wiley

Date: 11-1999

DOI: 10.1046/J.1365-2567.1999.00885.X

Abstract: In previous studies we have shown that histidine-rich glycoprotein (HRG), a relatively abundant plasma protein, can bind to immunoglobulin G (IgG) and inhibit the insolubilization of IgG-containing immune complexes (IC). It was of interest, therefore, to determine whether HRG can inhibit the formation of insoluble IC (IIC) resulting from the interaction of rheumatoid factor (RF) with human IgG-containing IC. Light scattering techniques were used to examine the effect of HRG on the formation of IIC between RF and IC containing human IgG according to three different models. In all three models physiological concentrations of HRG could block the formation of IIC induced by RF. Optical biosensor studies of the RF-IgG interaction also revealed that HRG can mask the epitopes on IgG recognized by RF. Additional studies examined whether HRG can solubilize already formed IIC and demonstrated that HRG can, in fact, partially solubilized IIC. These data indicate that HRG can regulate the formation of IIC induced by RF at three levels: namely by inhibiting the initial recognition of IgG containing IC by RF, by inhibiting the subsequent insolubilization of IgG containing IC by RF and by solubilizing already formed IIC. Collectively, these findings suggest that HRG may be an important inhibitor of the formation of pathogenic IC in diseases such as systemic lupus erythematosus and rheumatoid arthritis.

The yeast homolog of heme oxygenase-1 affords cellular antioxidant protection via the transcriptional regulation of known antioxidant genes

Publisher: Elsevier BV

Date: 2011

DOI: 10.1074/JBC.M110.187062

Chicken insulin-like growth factor-I: Amino acid sequence, radioimmunoassay, and plasma levels between strains and during growth

Publisher: Elsevier BV

Date: 09-1990

DOI: 10.1016/0016-6480(90)90076-X

Abstract: Insulin-like growth factor-I (IGF-I) has been purified from chicken serum and sequenced. The peptide has eight amino acid substitutions when compared with human IGF-I: serine26, leucine38, histidine39, histidine40, lysine41, glutamine50, isoleucine64, and proline67. Chicken IGF-I (cIGF-I) has been measured using a radioimmunoassay with a human IGF-I (hIGF-I) standard and an antibody raised against hIGF-I. In this assay the cross-reactivity of cIGF-I was approximately 50% with respect to hIGF-I and the cross-reactivity of chicken IGF-II was 1.7% with respect to chicken IGF-I. To determine whether binding proteins in chicken plasma can artifactually interfere with IGF-I measurements as they do in mammals, chicken plasma was fractionated by molecular sieve chromatography at acid pH. When the fractions corresponding to the binding protein region were included in the IGF-I radioimmunoassay, essentially no apparent IGF-I was detected, indicating that the binding proteins did not interfere. This result, together with the finding that IGF-I in acid-ethanol extracts of chicken plasma produced parallel dose-response curves to pure cIGF-I and hIGF-I, allows the reliable measurement of cIGF-I in such extracts. The concentrations of IGF-I in plasma from male birds increased two- to threefold between 1 and 7 weeks after hatching to achieve 30-45 ng/ml. Smaller increases were found in female chickens from a higher value at 1 week. No diurnal pattern of IGF-I levels could be detected. In 4-week-old birds, the plasma concentration of the peptide fell from nearly 40 to 15 ng/ml after 24 hr of starvation and to 9 ng/ml 20 hr later. These effects are very similar to those described for mammals and strongly suggest that the regulation of IGF-I is conserved during evolution, notwithstanding the lower plasma concentrations of the growth factor in chickens.

Inhibition of experimental allergic encephalomyelitis by the α-glucosidase inhibitor castanospermine

Publisher: Elsevier BV

Date: 03-1989

DOI: 10.1016/0022-510X(89)90047-6

Abstract: The alkaloid castanospermine is a potent inhibitor of oligosaccharide processing in vitro. Our recent findings indicating the importance of carbohydrate moieties in some critical step of the neuro-immunologic inflammatory process of allergic encephalomyelitis prompted us to investigate the effect of castanospermine on this disease process. The alkaloid inhibited passively induced allergic encephalomyelitis in a dose-dependent manner when administered continuously for 7 days beginning at the time of lymphocyte transfer. Although clinical disease was totally inhibited, treated animals did have inflammatory lesions in the central nervous system. These lesions were qualitatively different from those seen in untreated animals in that the inflammatory cells were tightly packed around the vessels and showed little migration into surrounding tissues. Castanospermine also effectively inhibited clinical disease in recipient animals which had had a previous episode of allergic encephalomyelitis. Castanospermine did not alter the disease when treatment was started after the onset of clinical symptoms.

H-2U: a new region at the D end of the murine MHC.

Publisher: Springer Science and Business Media LLC

Date: 1981

DOI: 10.1007/BF00350791

Abstract: Phylogenetics is the application of comparative studies of genetic sequences in order to infer evolutionary relationships among organisms. This tool can be used as a form of molecular epidemiology to enhance traditional population-level communicable disease surveillance. Phylogenetic study has resulted in new paradigms being created in the field of communicable diseases and this commentary aims to provide the reader with an explanation of how phylogenetics can be used in tracking infectious diseases. Special emphasis will be placed upon the application of phylogenetics as a tool to help elucidate HIV transmission patterns and the limitations to these methods when applied to forensic analysis. Understanding infectious disease epidemiology in order to prevent new transmissions is the sine qua non of public health. However, with increasing epidemiological resolution, there may be an associated potential loss of privacy to the in idual. It is within this context that we aim to promote the discussion on how to use phylogenetics to achieve important public health goals, while at the same time protecting the rights of the in idual.

Major histocompatibility gene complex (MHC)-coded determinants on antigen-specific suppressor factor for delayed-type hypersensitivity and surface phenotypes of cells producing the factor

Publisher: Wiley

Date: 04-1980

DOI: 10.1002/EJI.1830100415

Abstract: Antigen-specific suppressor factor for delayed-type hypersensitivity (DTH) to sheep red blood cells (SRBC) was obtained by incubating in vitro spleen cells from CBA mice (H-2k) injected intravenously 3 days previously with 1 x 10(9) SRBC. The suppressor factor was characterized for major histocompatibility gene complex (MHC)-coded antigenic determinants by passing the factor through immunosorbents coupled with appropriate alloantisera. The suppressor factor was absorbed by anti-H-2k, anti-Iak and anti-I-Jk immunosorbents but was not retained by anti-Ias, anti-I-Js, anti-I-Ak, anti-I-E/Ck or anti-H-2Kk immunosorbents. In addition, the factor bound to an immunosorbent coupled with rabbit antibodies against carbohydrate-defined Ia antigens. Furthermore, the suppressive activity that was absorbed was quantitatively recovered in the acid eluates from the immunosorbents. Treatment of the spleen cells with anti-Lyt-1.1 antiserum and complement completely abrogated their ability to elaborate the suppressor factor in vitro. In contrast, treatment with anti-Lyt-2.1 or anti-Iak antiserum and complement had no effect. Thus, it appears that the suppressor factor for DTH to SRBC bears I-J subregion-coded determinants, and its production is dependent on cells which have the Lyt-1+,2- and Ia- phenotype.

Interplay between chromatin remodeling and epigenetic changes during lineage-specific commitment to granzyme B expression

Publisher: The American Association of Immunologists

Date: 13-01-2010

DOI: 10.4049/JIMMUNOL.0990115

DNA-binding Proteins of Chick Embryo Lethal Orphan Virus: Lack of Complementation between Early Proteins of Avian and Human Adenoviruses

Publisher: Microbiology Society

Date: 10-1984

DOI: 10.1099/0022-1317-65-10-1817

Abstract: Chick cells infected by chick embryo lethal orphan (CELO) virus (fowl adenovirus type 1) contained four prominent virus-specific, structurally related DNA-binding proteins with mol. wt. of 74K, 64K, 56K, 52K, and two minor forms. The CELO virus DNA-binding proteins were phosphorylated, delayed-early nuclear proteins. CELO virus early proteins were expressed in BHK cells, but did not complement human adenovirus type 5 mutants with lesions E1A, E2A or E2B. Moreover, CELO virus DNA-binding proteins were not produced in 293 cells, which express human adenovirus E1 genes. These results suggest that activation of transcription by adenovirus E1A genes involves specific interactions between the E1A gene products and viral early promoters.

Lymphocytes express Ia antigens of foreign haplotype following treatment with neuraminidase.

Publisher: Springer Science and Business Media LLC

Date: 12-1981

DOI: 10.1007/BF01561647

Abstract: It has been recognized for a long time that gastric cancer behavior and outcomes might be different between patients living in Asian countries To evaluate the impact of race on survival outcomes of non-metastatic gastric cancer patients in the United States. This is a secondary analysis of a randomized controlled trial (CALGB 80101 study) that evaluated two adjuvant chemoradiotherapy schedules following resection of non-metastatic gastric cancer. Kaplan-Meier analysis and log-rank testing were utilized to explore the overall and disease-free survival differences according to the race of the patients. Univariate and multivariate Cox regression analyses were then used to explore factors affecting overall and disease-free survivals. A total of 546 patients were included in the current analysis. Of which, 73.8% have white race ( Asian American patients with non-metastatic gastric cancer have better overall and disease-free survival compared to other racial groups in the United States. Further preclinical and clinical research is needed to clarify the reasons behind this observation.

The accumulation of circulating histones on heparan sulphate in the capillary glycocalyx of the lungs

Publisher: Elsevier BV

Date: 07-2013

DOI: 10.1016/J.BIOMATERIALS.2013.03.091

Abstract: Recent findings on the role of circulating histone proteins in mediating acute lung injury prompted us to investigate whether there is a specific mechanism for accumulation of histones in the lungs. Binding sites for polycations are already known in the vasculature of the lungs, and we postulated that these could also be involved in histone accumulation, since histones have a high content of positively charged amino acids. Using a histone-coated colloid of a radiolabelled nanocomposite to track histone biodistribution with imaging techniques, it was found that histones bind avidly in the lungs of rabbits after intravenous injection. Blocking experiments with competing polycations in vivo characterised histone lung binding as dependent on a charge interaction with microvessel polyanions. Pretreatment of rabbits with a specific heparinase confirmed that the lung binding sites consist of heparan sulphate in the endothelial glycocalyx. A range of heparan sulphate analogues was accordingly shown to prevent histone accumulation in the lungs by neutralising histones in blood. These findings provide a rational basis for the design of polyanions that can prevent accumulation of cytotoxic histones in the lungs and thereby intervene at an early key step in the development of acute lung injury.

Secretion of Ia antigens by a subpopulation of T cells which are Ly-1+, Ly-2-, and Ia-.

Publisher: Rockefeller University Press

Date: 09-1976

DOI: 10.1084/JEM.144.3.847

Abstract: It was found that Ia antignes are rapidly secreted by a subpopulation of splenic T lymphocytes which are nonadherent and which express the surface phenotype Ly-1+, Ly-2-, and Ia-. Secretion of the Ia antigens was a metabolically active process which was inhibited by sodium azide and by Pactamycin, an inhibitor of protein synthesis.

Treatment of central nervous system inflammation with inhibitors of basement membrane degradation

Publisher: Wiley

Date: 02-1998

DOI: 10.1046/J.1440-1711.1998.00722.X

Abstract: Currently available anti-inflammatory drugs for the treatment of multiple sclerosis (MS) and other inflammatory diseases are generally inadequate, with disease progression not being arrested by the treatments and undesirable side effects posing problems. In response to these deficiencies our laboratories have, over the past 10 years, been developing novel drugs that interfere with the entry of leucocytes into inflammatory sites by inhibiting their passage through the subendothelial basement membrane (BM). This review initially summarizes evidence supporting the hypothesis that the subendothelial BM is a major barrier to the accumulation of leucocytes in inflammatory sites. An important point that has emerged is that breaching of the BM is probably a cooperative process, involving activation- and cytokine-induced degradative enzymes contributed by leucocytes, endothelial cells and platelets. The review then discusses the properties of three separate classes of anti-inflammatory compounds we have developed, namely sulfated polysaccharides/oligosaccharides, phosphosugars, and castanospermine (CS), which inhibit the passage of leukocytes through BM. Each drug type appears to prevent BM degradation by a different mechanism. Sulfated polysaccharides/oligosaccharides mediate their anti-inflammatory effect by inhibiting the endoglycosidase, heparanase, which plays a key role in the solubilization of BM by invading leucocytes. In fact, our studies have highlighted the heparanase enzyme as a major target for future drug development. Phosphosugars probably inhibit inflammation by displacing lysosomal enzymes, which are involved in BM degradation, from cell surface mannose 6-phosphate receptors. This mechanism of expressing degradative enzymes on the cell surface is particularly evident with activated T lymphocytes. On the other hand, CS interferes with appropriate targeting of lysosomal enzymes involved in BM degradation. For reasons which are still unclear, CS specifically inhibits BM degradation by endothelial cells, which results in a characteristic perivascular arrest of leucocytes in inflammatory sites. Overall, our studies have established that inhibitors of subendothelial BM degradation represent viable anti-inflammatory agents. It is hoped that future work will result in the development of a totally new class of highly effective, subtle and non-toxic anti-inflammatory drugs for the treatment of MS and other inflammatory diseases.

Inhibition of Passive Allergic Encephalomyelitis by Sulfated Polysaccharides

Publisher: Wiley

Date: 11-1988

DOI: 10.1111/J.1749-6632.1988.TB27166.X

Abstract: Exposure to disinfection by-products (DBPs) in drinking water has been associated with cancer risk. A recent study (Villanueva et al. 2007 Am J Epidemiol 165:148-156) found an increased bladder cancer risk among subjects attending swimming pools relative to those not attending. We evaluated adults who swam in chlorinated pools to determine whether exposure to DBPs in pool water is associated with biomarkers of genotoxicity. We collected blood, urine, and exhaled air s les from 49 nonsmoking adult volunteers before and after they swam for 40 min in an indoor chlorinated pool. We estimated associations between the concentrations of four trihalomethanes (THMs) in exhaled breath and changes in micronuclei (MN) and DNA damage (comet assay) in peripheral blood lymphocytes before and 1 hr after swimming urine mutagenicity (Ames assay) before and 2 hr after swimming and MN in exfoliated urothelial cells before and 2 weeks after swimming. We also estimated associations and interactions with polymorphisms in genes related to DNA repair or to DBP metabolism. After swimming, the total concentration of the four THMs in exhaled breath was seven times higher than before swimming. The change in the frequency of micronucleated lymphocytes after swimming increased in association with higher exhaled concentrations of the brominated THMs (p = 0.03 for bromodichloromethane, p = 0.05 for chlorodibromomethane, p = 0.01 for bromoform) but not chloroform. Swimming was not associated with DNA damage detectable by the comet assay. Urine mutagenicity increased significantly after swimming, in association with the higher concentration of exhaled bromoform (p = 0.004). We found no significant associations with changes in micronucleated urothelial cells. Our findings support potential genotoxic effects of exposure to DBPs from swimming pools. The positive health effects gained by swimming could be increased by reducing the potential health risks of pool water.

Carbohydrate recognition molecules on lymphocytes

Publisher: Portland Press Ltd.

Date: 05-1992

DOI: 10.1042/BST0200295

Abstract: In healthy controls (n = 8) living in shigella endemic areas, accumulation of interferon gamma (IFN gamma) in the epithelial lining was seen in the rectal tissues. At the single cell level, however, few or no IFN gamma protein producing cells or mRNA expressing cells were detected at that site indicating the involvement of the whole large intestine in the production of IFN gamma in controls. Persistent numbers of IFN gamma producing cells were detected in the rectum of patients with Shigella dysenteriae type 1 infection (n = 8) throughout the course of disease with a tendency to increase in the convalescent stage. A significantly increased extra cellular deposition of secreted IFN gamma in tissue was seen in convalescence when compared with the acute stage (p < 0.05). In addition, enzyme immunoassay showed increased stool concentration of IFN gamma in patients at the convalescent stage as well as in healthy controls. In situ hybridisation confirmed the results by showing increased frequency of IFN gamma mRNA containing cells at the late stage of the disease (p < 0.05). Extensive message for IFN gamma was evident in cells in the lamina propria with no detectable transcripts in the surface epithelium. A colocalisation of IFN gamma with the IFN gamma receptor expression, predominantly found in the epithelial lining was detected by immunohistochemistry. Semiquantitative evaluation by computerised image analysis showed a gradual increased expression of IFN gamma and its corresponding receptor in the convalescent stage of shigellosis. This suggested progressive entrapment and binding of IFN gamma to its specific receptor at the local site. The enhanced surface expression of IFN gamma receptor evident at the convalescent stage of shigellosis was comparable to the constitutive level of expression in the healthy subjects. Thus, immunity to shigellosis correlated to up-regulation of IFN gamma production and expression of IFN gamma receptor.

Cleavage of bacterial flagellin with proteolytic enzymes. II. Induction of enhanced delayed-type hypersensitivity to flagellin by peptic fragments of the protein

Publisher: Elsevier BV

Date: 07-1972

DOI: 10.1016/0008-8749(72)90030-5

Abstract: For the application of compressive sensing to parallel MRI, Poisson disk s ling (PDS) has been shown to generate superior results compared with random s ling methods. However, due to its limited flexibility to incorporate additional constraints, PDS is not readily extendible to dynamic applications. Here, we propose and validate a pseudo-random s ling technique that allows incorporating constraints specific to dynamic imaging. The proposed s ling scheme, called variable density incoherent spatiotemporal acquisition (VISTA), is based on constrained minimization of Riesz energy on a spatiotemporal grid. Data from both a digital phantom and real-time cine were used to compare VISTA with uniform interleaved s ling (UIS) and variable density random s ling (VRS). The image quality was assessed qualitatively and quantitatively. VISTA improved the trade-off between noise and sharpness. Also, VISTA produced diagnostic quality images at an acceleration rate of 15, whereas UIS and VRS images degraded below the diagnostic threshold at lower acceleration rates. VISTA generates spatiotemporal s ling patterns with high levels of uniformity and incoherence, while maintaining a constant temporal resolution. Using a small pilot study, VISTA was shown to produce diagnostic quality images at acceleration rates up to 15.

Leptin signals via TGFB1 to promote metastatic potential and stemness in breast cancer

Publisher: Public Library of Science (PLoS)

Date: 25-05-2017

DOI: 10.1371/JOURNAL.PONE.0178454

Evidence that carboxyl-reduced heparin fails to potentiate acidic fibroblast growth factor activity due to an inability to interact with cell surface heparin receptors

Publisher: Elsevier BV

Date: 03-1995

DOI: 10.1006/EXCR.1995.1072

Abstract: Recently we reported that carboxyl-reduced heparin (CR-heparin), despite binding acidic fibroblast growth factor (aFGF) as effectively as native heparin, was much less potent at augmenting aFGF-induced mitogenesis. This paper describes experiments which examined this phenomenon in more detail in the hope that it would shed light on the mechanism by which heparin potentiates aFGF activity. Initial studies confirmed that heparin, with 60% of its carboxyl groups reduced, although binding aFGF with the same affinity as native heparin (Kd 35 +/- 5 nM), was a poor potentiator of aFGF-induced mitogenic activity. Proteolysis protection experiments also revealed that CR-heparin was as effective as native heparin at protecting aFGF from proteolytic degradation. In contrast, CR-heparin was considerably less effective than native heparin at enhancing the binding of aFGF to the fibroblast growth factor receptor (FGFR) on 3T3 cells. Furthermore, CR-heparin only bound to a subset (approximately 1/3) of heparin receptors on 3T3 cells. Based on these data, it is proposed that CR-heparin is less efficient than heparin at facilitating the formation of a quaternary complex among aFGF, the FGFR, and cell surface heparin receptors.

Phase I trial of Lipovaxin-MM, a novel dendritic cell-targeted liposomal vaccine for malignant melanoma

Publisher: Springer Science and Business Media LLC

Date: 16-07-2018

DOI: 10.1007/S00262-018-2207-Z

Abstract: In this phase I study using a 3 + 3 dose escalation design, the safety, dose-limiting toxicity (DLT), immunogenicity and efficacy of intravenous Lipovaxin-MM-a multi-component dendritic cell-targeted liposomal vaccine against metastatic melanoma-was investigated. Twelve subjects with metastatic cutaneous melanoma were recruited in three cohorts. Patients in Cohort A (n = 3) and Cohort B (n = 3) received three doses of 0.1 and 1 mL of Lipovaxin-MM, respectively, every 4 weeks. Patients in Cohort C (n = 6) received four doses of 3 mL vaccine weekly. Immunologic assessments of peripheral blood were made at regular intervals and included leukocyte subsets, cytokine levels, and Lipovaxin-MM-specific T-cell and antibody reactivities. Tumor responses were assessed by RECIST v1.0 at screening, then 8 weekly in Cohorts A and B and 6 weekly in Cohort C. Of a total of 94 adverse events (AEs) reported in ten subjects, 43 AEs in six subjects were considered to be possibly or probably vaccine-related. Most (95%) vaccine-related AEs were grade 1 or 2, two (5%) grade 3 vaccine-related AEs of anemia and lethargy were recorded, and higher grade AEs and DLTs were not observed. No consistent evidence of vaccine-specific humoral or cellular immune responses was found in post-immunization blood s les. One patient had a partial response, two patients had stable disease, and the remaining patients had progressive disease. Lipovaxin-MM was well tolerated and without clinically significant toxicity. Immunogenicity of Lipovaxin-MM was not detected. Partial response and stable disease were observed in one and two patients, respectively.

Histidine-rich glycoprotein functions cooperatively with cell surface heparan sulfate on phagocytes to promote necrotic cell uptake

Publisher: Oxford University Press (OUP)

Date: 23-06-2010

DOI: 10.1189/JLB.0210087

Abstract: HRG enhances the phagocytosis of necrotic cells via a heparan sulfate-dependent pathway that is inhibitable by heparin. Dying cells, such as apoptotic and necrotic cells, are cleared rapidly from the site of cell death to prevent the exposure of intracellular antigenic and immunostimulatory molecules that may cause tissue injury or facilitate the development of autoimmune diseases. For the immune system to recognize and remove dying cells efficiently, professional phagocytes use a variety of mechanisms that distinguish healthy cells from dying cells. HRG, a relatively abundant heparin/HS-binding protein in human plasma, has been shown recently to tether IgG specifically to necrotic cells and aid the phagocytic uptake of necrotic cells via a FcγRI-dependent pathway. In this study, we provide direct evidence that HRG can function cooperatively with cell surface HS on the monocytic cell line THP-1 to promote necrotic cell removal. In addition, we found that the presence of heparin can markedly inhibit HRG-enhanced necrotic cell clearance by THP-1 cells, possibly by blocking the ability of HRG to interact with necrotic cells as well as THP-1 cells. Thus, these data suggest that HRG can aid the phagocytosis of necrotic cells via a HS-dependent pathway, and this process can be regulated by the presence of certain HRG ligands, such as heparin.

Dramatic regulation of heparanase activity and angiogenesis gene expression in synovium from patients with rheumatoid arthritis

Publisher: Wiley

Date: 31-05-2008

DOI: 10.1002/ART.23489

Abstract: Although heparanase is recognized as a proangiogenic factor, the involvement of heparanase in rheumatoid arthritis (RA) is unclear. In this study, we assessed heparanase activity in synovial fluid (SF) and synovial tissue (ST) from patients with RA or osteoarthritis (OA), and analyzed the expression of angiogenic pathway-focused genes in ST from RA and OA patients. SF and ST were obtained from the knees of patients with either RA or OA and from asymptomatic donors with no documented history of degenerative or inflammatory joint diseases. Heparanase activity was determined by an enzymatic assay using a radiolabeled substrate, and the presence of heparanase in ST was demonstrated by Western blotting. The expression of angiogenesis genes, including heparanase, in ST was analyzed by real-time quantitative polymerase chain reaction. Heparanase activity was dramatically higher (>100-fold) in SF and ST from RA patients than in SF and ST from OA patients and asymptomatic donors. Active heparanase enzyme was detected and heparanase messenger RNA was up-regulated in ST from RA patients. We also found that angiogenesis gene expression was significantly regulated in RA synovium, and was correlated with heparanase activity. These findings are novel and contribute to our understanding of joint destruction in RA, suggesting that heparanase may be a reliable prognostic factor for RA progression and an attractive target for the treatment of RA.

T cell receptor sharing by cytotoxic T lymphocytes facilitates efficient virus control

Publisher: Proceedings of the National Academy of Sciences

Date: 09-2009

DOI: 10.1073/PNAS.0906554106

Abstract: A remarkable feature of the adaptive immune system is the speed at which small numbers of antigen-specific lymphocytes can mediate a successful immune response. Rapid expansion of T and B lymphocyte clones that have receptors specific for a particular antigen is one of the primary means by which a swift response is generated. Although much of this clonal expansion is caused by the ision of antigen-specific cells, here we demonstrate an additional mechanism by which the pool of effector T cells against a viral infection can quickly enlarge. Our data show that virus-specific CD8 + cytotoxic T lymphocytes (CTL) can transfer their T cell receptors (TCR) to recipient CTL of an unrelated specificity that, as a consequence, gain the antigen specificity of the donor T cell. This process occurs within minutes via membrane exchange and results in the recipient CTL acquiring the ability to recognize and eliminate cells targeted by the donor TCR, while still retaining the antigen specificity of its own TCR. Such receptor sharing allows rapid, proliferation-independent expansion of virus-specific T cell clones of low frequency and plays a highly significant antiviral role that can protect the host from an otherwise lethal infection.

Immune response to chemically modified flagellin. IV. Further studies on the relationship between humoral and cell-mediated immunity

Publisher: Elsevier BV

Date: 1973

DOI: 10.1016/0008-8749(73)90007-5

Transfer of cell-mediated immunity with cell-free leukocyte extracts. III. Demonstration of Ia antigens in the specific component.

Publisher: Elsevier BV

Date: 12-1980

DOI: 10.1016/0008-8749(80)90119-7

Abstract: To explore palliative care and oncology clinicians' perspectives on current challenges and facilitating factors in meeting the spiritual needs of patients with lung cancer and family caregivers. This study was conducted in preparation for a community-based lung cancer palliative care intervention. 19 oncology and palliative care clinicians in three outpatient Kaiser Permanente sites in southern California. This multisite qualitative study used focus group and key informant interviews. Data were analyzed using content analysis methodology, and a team approach was used to validate findings. Clinicians described facilitating factors (interprofessional team support, assessment of spiritual needs, clinician-provided spiritual support, and provision of culturally respectful spiritual care) and challenges (related to providing culturally respectful spiritual care by respecting the patients' spiritual and cultural beliefs in an open way and in advocating for the patients' wishes) they encountered when addressing patient and caregiver spiritual needs. This study demonstrated the need to provide nurses with practical tools, education, and a supportive environment to address patients' and family caregivers' spiritual concerns.

Regulation of histidine-rich glycoprotein (HRG) function via plasmin-mediated proteolytic cleavage

Publisher: Portland Press Ltd.

Date: 23-10-2009

DOI: 10.1042/BJ20090794

Abstract: The plasminogen lasmin system is involved in a variety of normal physiological and pathological processes, including tissue remodelling, angiogenesis and tumour metastasis. Plasminogen activators and receptors for plasminogen lasminogen activators are essential for the processing of plasminogen to form the active serine protease plasmin. Plasmin can in turn positively or negatively regulate further plasminogen activation via plasminmediated cleavage of receptors and activators. HRG (histidine-rich glycoprotein), a relatively abundant (approx. 100–150 μg/ml) plasma glycoprotein, has a multi-domain structure that can interact with many ligands, including Zn2+, heparin, HS (heparan sulfate) and plasminogen. HRG has been shown to function as an adaptor molecule to tether plasminogen to GAG (glycosaminoglycan)-bearing surfaces and to regulate plasminogen activation via various mechanisms. As HRG itself is sensitive to plasmin cleavage, the present study examines in detail the cleavage of human HRG by plasmin and the effect of this cleavage on various functions of HRG. HRG fragments, generated by plasmin cleavage, are held together by disulfide linkages and are not released from the molecule under non-reducing conditions. Plasmin-mediated cleavage partially inhibited HRG binding to cell surface HS, but enhanced HRG binding to necrotic cells and to plasminogen. However, both intact and plasmin-cleaved HRG enhanced the binding of plasminogen to heparin-coated surfaces to a similar extent. Furthermore, the presence of heparin, Zn2+ or acidic pH was found to protect HRG from plasmin cleavage. Thus proteolytic cleavage of HRG by plasmin may provide a feedback mechanism to regulate the effects of HRG on the plasminogen lasmin system and other functions of HRG.

Antiangiogenic platinum through glycan targeting

Publisher: Royal Society of Chemistry (RSC)

Date: 2017

DOI: 10.1039/C6SC02515C

Abstract: The high affinity of highly charged polynuclear platinum complexes for glycans such as heparan sulfate results in modulation of the biomolecule signaling functions leading to inhibition of angiogenesis.

Monoclonal antibody detection of two classes of H-2Kk molecules.

Publisher: Elsevier BV

Date: 08-1981

DOI: 10.1016/0161-5890(81)90063-8

Cloning of mammalian heparanase, an important enzyme in tumor invasion and metastasis

Publisher: Springer Science and Business Media LLC

Date: 07-1999

DOI: 10.1038/10525

Abstract: The endoglycosidase heparanase is an important in the degradation of the extracellular matrix by invading cells, notably metastatic tumor cells and migrating leukocytes. Here we report the cDNA sequence of the human platelet enzyme, which encodes a unique protein of 543 amino acids, and the identification of highly homologous sequences in activated mouse T cells and in a highly metastatic rat adenocarcinoma. Furthermore, the expression of heparanase mRNA in rat tumor cells correlates with their metastatic potential. Exhaustive studies have shown only one heparanase sequence, consistent with the idea that this enzyme is the dominant endoglucuronidase in mammalian tissues.

Acidic and basic fibroblast growth factor bind with differing affinity to the same heparan sulfate proteoglycan on BALB/c 3T3 cells: Implications for potentiation of growth factor action by heparin

Publisher: Wiley

Date: 05-1995

DOI: 10.1002/JCB.240580103

Abstract: Heparan sulfate proteoglycans on the cell surface act as low affinity binding sites for acidic and basic fibroblast growth factor (FGF) [Moscatelli (1987): J Cell Physiol 131:123-130] and play an important role in the interaction of FGF with the FGF receptor (FGFR). In this study, several aspects of the interaction of FGFs with cell surface heparan sulfate proteoglycans were examined. Reciprocal cross blocking studies demonstrated that acidic FGF (aFGF) and basic FGF (bFGF) bind to identical or closely associated heparan sulfate motifs on BALB/c 3T3 cell surface heparan sulfate proteoglycans. However, the binding affinity of the two growth factors for these heparan sulfate proteoglycans differs considerably, competition binding data indicating that aFGF has a 4.7-fold lower affinity than bFGF for 3T3 heparan sulfate proteoglycan. Subsequent studies of dissociation kinetics demonstrated that bFGF dissociates from the FGFR at least 10-fold slower than aFGF, whereas, following removal of cell surface heparan sulfate proteoglycans by heparinase treatment, the dissociation rate of both FGFs is similar and rapid. These results support the concept that cell surface heparan sulfate proteoglycans stabilize the interaction of FGF with FGFR, possibly by the formation of a ternary complex.

Preferential induction of cell-mediated immunity by chemically modified sheep erythrocytes

Publisher: Wiley

Date: 04-1972

DOI: 10.1002/EJI.1830020210

Abstract: Selection bias occurs when recruiters selectively enrol patients into the trial based on what the next treatment allocation is likely to be. This can occur even if appropriate allocation concealment is used if recruiters can guess the next treatment assignment with some degree of accuracy. This typically occurs in unblinded trials when restricted randomisation is implemented to force the number of patients in each arm or within each centre to be the same. Several methods to reduce the risk of selection bias have been suggested however, it is unclear how often these techniques are used in practice. We performed a review of published trials which were not blinded to assess whether they utilised methods for reducing the risk of selection bias. We assessed the following techniques: (a) blinding of recruiters (b) use of simple randomisation (c) avoidance of stratification by site when restricted randomisation is used (d) avoidance of permuted blocks if stratification by site is used and (e) incorporation of prognostic covariates into the randomisation procedure when restricted randomisation is used. We included parallel group, in idually randomised phase III trials published in four general medical journals (BMJ, Journal of the American Medical Association, The Lancet, and New England Journal of Medicine) in 2010. We identified 152 eligible trials. Most trials (98%) provided no information on whether recruiters were blind to previous treatment allocations. Only 3% of trials used simple randomisation 63% used some form of restricted randomisation, and 35% did not state the method of randomisation. Overall, 44% of trials were stratified by site of recruitment 27% were not, and 29% did not report this information. Most trials that did stratify by site of recruitment used permuted blocks (58%), and only 15% reported using random block sizes. Many trials that used restricted randomisation also included prognostic covariates in the randomisation procedure (56%). The risk of selection bias could not be ascertained for most trials due to poor reporting. Many trials which did provide details on the randomisation procedure were at risk of selection bias due to a poorly chosen randomisation methods. Techniques to reduce the risk of selection bias should be more widely implemented.

Immunology and cell biology turns 90

Publisher: Wiley

Date: 26-11-2014

DOI: 10.1038/ICB.2013.89

Nonimmune lymphocyte-macrophage interaction

Publisher: Elsevier BV

Date: 05-1985

DOI: 10.1016/0008-8749(85)90008-5

Abstract: Previous studies have demonstrated a spontaneous, nonimmune interaction between lymphocytes and macrophages. This paper describes an automated colorimetric assay based on the dye, rose bengal, to quantify this interaction. The procedure entails allowing lymphocytes to adhere to preformed macrophage monolayers in the wells of microplates and then staining bound lymphocytes with rose bengal. Dye uptake and the consequent number of lymphocytes bound were quantified using an automated spectrophotometer developed for reading microplates. This procedure was used to confirm and extend the basic parameters of the system. The interaction was found to be temperature dependent but the kinetics and percentage of cells binding varied with the source of lymphocytes. However, all lymphocyte populations tested, namely, mature and immature thymocytes, T and B lymphocytes, and a range of thymoma cell lines, bound to macrophages. Furthermore, all macrophage populations examined had the ability to bind lymphocytes. The interaction also showed no strain specificity and generally lacked species specificity. It is proposed that the interaction is a highly dynamic process that enables lymphocytes to scan the surface of macrophages for self and/or foreign antigens.

Heparanase: Historical Aspects and Future Perspectives

Publisher: Springer International Publishing

Date: 2020

DOI: 10.1007/978-3-030-34521-1_3

Engrafting costimulator molecules onto tumor cell surfaces with chelator lipids: A potentially convenient approach in cancer vaccine development

Publisher: The American Association of Immunologists

Date: 03-2000

DOI: 10.4049/JIMMUNOL.164.5.2433

Abstract: The genetic modification of cells to develop cell-based vaccines and to modulate immune responses in vivo can be risky and inconvenient to perform in clinical situations. A novel chelator lipid, nitrilotriacetic acid di-tetradecylamine (NTA-DTDA) that, via the NTA group has high affinity for 6His peptide, was used to directly anchor recombinant forms of T cell costimulatory molecules containing a C-terminal 6-His sequence onto tumor cell surfaces. Initial experiments using murine P815 tumor cells established the optimum conditions for incorporating NTA-DTDA onto the membranes of cells. P815 cells with incorporated NTA-DTDAbound hexahistidine-(6His)-tagged forms of the extracellular domains of murine B7.1 and CD40 (B7.1-6H and CD40-6H) at very high levels (fluorescence 200–300-fold above background), and both proteins could be anchored onto the cells simultaneously. Significant loss of the anchored or “engrafted” protein occurred through membrane internalization following culture of the cells under physiological conditions, but P815 cells with engrafted B7.1-6H and/or CD40-6H stimulated the proliferation of allogenic and syngeneic splenic T cells in vitro, and generated cytotoxic T cells when used as vaccines in syngeneic animals. Furthermore, the immunization of syngeneic mice with P815 cells engrafted with B7.1-6H or with B7.1-6H and CD40-6H induced protection against challenge with the native P815 tumor. The results indicate that the use of chelator lipids like NTD-DTDA to engraft costimulatory and/or other molecules onto cell membranes could provide a convenient alternative to transfection in the development of cell-based vaccines and for modulation of immune function.

Immune response to chemically modified flagellin. III. Enhanced cell-mediated immunity during high and low zone antibody tolerance to flagellin

Publisher: Rockefeller University Press

Date: 02-1972

DOI: 10.1084/JEM.135.2.298

Abstract: High and low zone antibody tolerance to bacterial flagellin can be induced in adult strain W Wistar rats by multiple injections of a cyanogen bromide (CNBr) digest of flagellin at two widely spaced dose levels. Intermediate doses of the CNBr digest produce enhanced antibody titers to flagellin rather than antibody tolerance. Studies reported in this paper revealed that both high and low zone antibody tolerance to flagellin were accompanied by heightened levels of delayed-type hypersensitivity. Conversely, when enhancement of the antibody response occurred, suppression of delayed hypersensitivity was observed. This inverse relationship between humoral and cell-mediated immunity was very striking in strain W Wistar rats but was not quite so clear-cut in another strain of Wistar rats (strain J). Strain J rats were resistant to the induction of antibody tolerance and gave higher immunological responses to flagellin than strain W animals. In addition, it was observed that, in contrast to adult tolerance, administration of the CNBr digest to neonatal rats induced complete tolerance at the level of both humoral and cell-mediated immunity. These findings were discussed in the light of earlier studies with flagellin and provide further evidence for a previously described hypothesis.

Heparanase and Type 1 Diabetes

Publisher: Springer International Publishing

Date: 2020

DOI: 10.1007/978-3-030-34521-1_24

Automated colorimetric assay for T cell cytotoxicity

Publisher: Elsevier BV

Date: 03-1983

DOI: 10.1016/0022-1759(83)90277-6

Abstract: A colorimetric method has been developed for detecting the lysis of target cells by cytotoxic T lymphocytes (Tc). The method entails incubating Tc cells with thioglycollate-induced macrophage targets and estimating macrophage survival at the end of the assay by staining viable macrophages with the dye neutral red. The method is substantially more sensitive than the 51Cr release assay and can be used to detect alloreactive Tc cells and H-2-restricted Tc cells against viruses, haptens and minor-H antigens. Furthermore, the assay is applicable to limit dilution analysis of Tc cell precursors. The method is cheap, avoids radioactive materials and by measuring optical densities with automated spectrophotometers developed for microELISA systems, results can be obtained 50-100 times faster than with the radioactive procedure.

A one-pot synthesis and biological activity of ageladine A and analogues

Publisher: American Chemical Society (ACS)

Date: 17-03-2011

DOI: 10.1021/JM200039M

Abstract: A one-pot synthesis of ageladine A and analogues is reported. The key Pictet-Spengler reaction between 2-aminohistamine and aryl aldehydes has been successfully utilized for the synthesis of the natural product and 14 analogues. These compounds were screened for their matrix metalloprotease (MMP) and kinase inhibition to develop the first structure-activity relationship of ageladine A analogues. One compound, which showed significant kinase activity but little MMP inhibitory activity, was found to be highly active in an antiangiogenic screen, suggesting that the angiogenic activity of ageladine A is not associated with MMP inhibition but rather kinase inhibitory activity. Cytotoxicity was excluded as a mode of action by the assay of ageladine A and an analogue against 60 human cell lines.

Targeted liposomal delivery of TLR9 ligands activates spontaneous antitumor immunity in an autochthonous cancer model

Publisher: The American Association of Immunologists

Date: 15-07-2009

DOI: 10.4049/JIMMUNOL.0900736

Abstract: Accessibility of tumors for highly effective local treatment represents a major challenge for anticancer therapy. Immunostimulatory oligodeoxynucleotides (ODN) with CpG motifs are ligands of TLR9, which prime spontaneous antitumor immunity, but are less effective when applied systemically. We therefore developed a liposome-based agent for selective delivery of CpG-ODN into the tumor environment. A peptide that specifically targets angiogenic endothelial cells in a transgenic tumor model for islet cell carcinogenesis was engrafted into CpG-ODN containing liposomes. Intravenous injection of these liposomes resulted in specific accumulation around tumor vessels, increased uptake by tumor-resident macrophages, and retention over time. In contrast, nontargeted liposomes did not localize to the tumor vasculature. Consequently, only vascular targeting of CpG-ODN liposomes provoked a marked inflammatory response at vessel walls with enhanced CD8+ and CD4+ T cell infiltration and, importantly, activation of spontaneous, tumor-specific cytotoxicity. In a therapeutic setting, 40% of tumor-bearing, transgenic mice survived beyond week 45 after systemic administration of vascular-directed CpG-ODN liposomes. In contrast, control mice survived up to 30 wk. Therapeutic efficacy was further improved by increasing the frequency of tumor-specific effector cells through adoptive transfers. NK cells and CD8+ T cells were major effectors which induced tumor cell death and acted in conjunction with antivascular effects. Thus, tumor homing with CpG-ODN-loaded liposomes is as potent as direct injection of free CpG-ODN and has the potential to overcome some major limitations of conventional CpG-ODN monotherapy.

Isolation, tissue distribution, and chromosomal localization of a novel testis-specific human four-transmembrane gene related to CD20 and FcεRI-β

Publisher: Elsevier BV

Date: 2001

DOI: 10.1006/BBRC.2000.4088

Presence of receptors for self MHC antigens on non-T cells.

Publisher: Elsevier BV

Date: 09-1981

DOI: 10.1016/0167-5699(81)90058-X

Whole-exome sequencing in evaluation of patients with venous thromboembolism

Publisher: American Society of Hematology

Date: 29-06-2017

DOI: 10.1182/BLOODADVANCES.2017005249

Abstract: Using WES, we designed an extended thrombophilia panel consisting of 55 genes of significance to thrombosis. The extended thrombophilia panel identified multiple novel genetic variants with predicted roles in thrombosis or thrombophilia.

Signal minus 1: A key factor in immunological tolerance to tissue-specific self antigens?

Publisher: Wiley

Date: 06-1996

DOI: 10.1038/ICB.1996.49

Abstract: Recent data suggest that many autoreactive T cells, particularly to tissue-specific self antigens, can escape thymic deletion. The current dogma is that these autoreactive T cells are silenced by the failure of most tissues to provide co-stimulation (signal 2), antigen alone (signal 1) inducing T cell unresponsiveness. However, I propose that activation of autoreactive T cells frequently occurs but autodestruction by effector T cells is tightly regulated. This phenomenon is most evident with lymph node metastasizing tumour cells where the regional lymph node can mount a vigorous response to the invading tumour cells but tumour growth is unimpaired. I suggest that autodestruction is prevented by inhibitory receptors on T cells which recognize class I MHC structures on target cells. These receptors, which I propose deliver 'signal minus 1' to T cells, were recently described on NK cells and a subpopulation of peripheral T cells. They are also strikingly similar to a family of anti-self receptors that my laboratory described on murine T and B cells 15 years ago. In the 'signal minus 1' model, antigen-activated T cells acquire the inhibitory receptors when they become co-stimulation independent and gain the ability to exit lymphoid organs and enter non-lymphoid tissues. Thus, if autoreactive effector T cells encounter autoantigen in tissues they are functionally silenced by inhibitory receptor engagement and signal minus 1 delivery. In contrast, I propose that in response to intracellular infections, cells down-regulate expression of their ligands for inhibitory receptors. Such a model allows infected cells to be selectively eliminated by effector T cells. If correct, the model predicts that effector T cells, whether foreign-antigen- or autoantigen-specific, can selectively respond to infected cells. This apparent 'usefulness' of autoreactive T cells may explain their observed persistence even after an encounter with autoantigen. It is also suggested that signal minus 1 may silence autoreactive B cells specific for tissue-specific cell surface antigens and lack of signal minus 1 may partially explain the vigorous T cell response to allogeneic MHC. Finally, it is hypothesized that, in evolutionary terms, inhibition of autodestruction by the recognition of a 'self marker' and delivery of signal minus 1 is an ancient process which probably emerged in early metazoans.

The human Ia system: Definition and characterization by xenogeneic antisera

Publisher: Springer Science and Business Media LLC

Date: 12-1979

DOI: 10.1007/BF01561429

Use of the intracellular fluorescent dye CFSE to monitor lymphocyte migration and proliferation

Publisher: Wiley

Date: 02-2009

DOI: 10.1002/0471142735.IM0409S84

Abstract: The stable incorporation of the intracellular fluorescent dye 5‐(and ‐6)‐carboxyfluorescein diacetate succinimidyl ester (CFSE) into cells provides a powerful tool to monitor cell migration, and to quantify cell ision, because of the sequential decrease in fluorescent labeling in daughter cells. CFSE‐labeled lymphocytes have been used to analyze the relationship between cell ision and differentiation of cell function, and cell proliferation versus apoptosis, both in vivo and in vitro, and have allowed analysis of the site of response to antigens in vivo. Curr. Protoc. Immunol . 84:4.9.1‐4.9.13. © 2009 by John Wiley & Sons, Inc.

Comment on: Korpos et al. The peri-islet basement membrane, a barrier to infiltrating leukocytes in type 1 diabetes in mouse and human. Diabetes 2013;62:531-542

Publisher: American Diabetes Association

Date: 17-07-2013

DOI: 10.2337/DB13-0470

Direct visualization of T lymphocytes bearing Ia antigens controlled by the I-J subregion

Publisher: Rockefeller University Press

Date: 08-1977

DOI: 10.1084/JEM.146.2.332

Abstract: Previous studies have demonstrated that I-J-subregion-controlled Ia antigens are only expressed on a small subpopulation of peripheral T lymphocytes which includes the suppressor T cells of antibody responses (6). This subpopulation of T cells cannot be detected by conventional dye-exclusion cytotoxicity tests. A sensitive rosetting procedure therefore was developed for detecting the binding of anti-Ia antibodies to T lymphocytes. This assay system, unlike the complement lysis technique, has a low background and since it represents a direct binding assay could detect noncomplement-fixing antibodies in the antisera. Anti-Ia sera were absorbed with B cells and using the rosetting procedure in genetic mapping studies the remaining antibodies were found to be directed against I-J-subregion-controlled determinants. These determinants were shown to be highly haplotype specific for H-2(k) and H-2(s) and appeared to be exclusively expressed on Ly-l.l(-), Ly2.1(+), T lymphocytes, at least some of which were suppressor T cells. Lymphoid organs differed in their content of anti-I-J-reactive cells, the hierarchy being spleen, lymph node more than thymus, bone marrow. In contrast, on a T-cell basis, a high proportion (35 percent) of the T cells in bone marrow reacted with anti-I-J antibodies, a substantial proportion (13 percent) of T cells from spleen were reactive, whereas the lymph node and thymus T-cell populations contained only a small proportion of positive cells (1-4 percent).

Potential contrasting effects of platelets on the migration and invasion of sarcomas versus carcinomas

Publisher: Informa UK Limited

Date: 14-07-2021

DOI: 10.1080/09537104.2020.1792433

Murine histidine-rich glycoprotein: Cloning, characterization and cellular origin

Publisher: Wiley

Date: 06-2000

DOI: 10.1046/J.1440-1711.2000.00940.X

Abstract: Histidine-rich glycoprotein (HRG) is a plasma protein of vertebrates that has been implicated in the regulation of several important biological functions, including the immune response and blood clotting. In the present study, we have isolated and determined the sequence of the cDNAs for both mouse and rat HRG. The deduced amino acid sequences of mouse and rat HRG are 525 and 510 amino acids, respectively, and they show the same three-domain structure that has been predicted for human HRG, with which they share high amino acid identity. Northern blot analysis indicates that the mouse HRG mRNA is 1.7 kb and is localized specifically to the liver. It has been suggested, somewhat controversially, that some immune cells, such as monocytes and megakaryocytes, also synthesize HRG. Reverse transcriptase-polymerase chain reaction analysis has failed to show any HRG mRNA in immune tissues of the mouse, including the spleen, thymus, lymph node, bone marrow and peripheral blood leucocytes. These data suggest that HRG expression by immune cells is due to the acquisition of plasma HRG derived from the liver. Finally, genomic Southern blot analysis of the mouse HRG gene suggests that it is a single copy gene.

Differential binding of histidine-rich glycoprotein (HRG) to human IgG subclasses and IgG molecules containing κ and λ light chains

Publisher: Elsevier BV

Date: 10-1999

DOI: 10.1074/JBC.274.42.29633

Platelets, selectins, and the control of tumor metastasis

Publisher: Elsevier BV

Date: 06-2014

DOI: 10.1053/J.SEMINONCOL.2014.04.003

Abstract: The significant role of platelets and P-selectin in assisting tumor cell metastasis to the lungs has been frequently reported and reviewed. However, evidence recently has come to light on other pro-metastatic mechanisms of platelets beyond that of tumor cell protection from immune cell attack and aiding extravasation, such as promoting epithelial to mesenchymal transition in tumor cells and conveying signals from the primary tumor to distant tissues that optimize conditions for metastasis. Moreover, the role of platelets and selectins in hematogenous metastasis to frequently targeted organs other than the lungs has been less well examined. This review aims to summarize the literature on the roles of platelets in all stages of the metastatic process and to examine the participation of platelets and selectins in hematogenous metastasis to the lungs, liver, bone, and brain. In the light of the available evidence, potential therapeutic avenues for the control of metastasis are also discussed.

Experimental melanin-induced uveitis in the Fischer 344 rat is inhibited by anti-CD4 monoclonal antibody, but not by mannose-6-phosphate

Publisher: Oxford University Press (OUP)

Date: 1999

DOI: 10.1046/J.1365-2249.1999.00765.X

Abstract: Experimental melanin-induced uveitis (EMIU) is a rodent model of acute anterior uveitis which was described in 1993. We investigated strain susceptibility, and age and gender characteristics of the model, undertook histological and immunohistochemical studies to investigate underlying cellular mechanisms, and examined several treatment options. Rats were immunized with bovine ocular melanin (250 μg), and disease was followed by slit l examination. Lewis, Fischer 344 and Porton rats were found to be susceptible to EMIU, whereas Wistar-Furth, DA, and Hooded Wistar strains were resistant. EMIU was neither age- nor gender-dependent. In Fischer 344 rats, EMIU was characterized clinically by florid anterior segment inflammation. Histopathological findings included infiltration of ciliary body and iris with mononuclear cells and neutrophils. Both CD4+ and CD8+ T lymphocytes were prominent. Rats were then treated with intraperitoneal injections of anti-CD4, anti-CD8 or irrelevant isotype-matched MoAb on days −3, 0, 3, 6 and 9 with respect to melanin immunization. Incidence of uveitis was significantly reduced in rats treated with a non-depleting cocktail of anti-CD4 MoAbs (P = 0.007), whereas a depleting anti-CD8 antibody had no effect on the disease. Mannose-6-phosphate inhibits lymphocyte migration in some models of T cell-mediated inflammation. This simple sugar was administered to additional rats via intraperitoneal osmotic pumps for 14 days following disease induction, but did not influence the uveitis. We conclude that EMIU is controlled by CD4+ T cells, and disease may be abrogated by treatment with anti-CD4 MoAbs.

The use of fluorescent target arrays for assessment of T cell responses in vivo

Publisher: MyJove Corporation

Date: 19-06-2014

DOI: 10.3791/51627

Latex bead rosetting method for cell surface antigens

Publisher: Elsevier BV

Date: 09-1982

DOI: 10.1016/0022-1759(82)90183-1

Abstract: Polystyrene latex beads coupled with anti-Ig can be used to detect the binding of specific antibodies to target cells. The optimal conditions for coupling anti-Ig to beads are described. This method is as sensitive as red cell rosetting and has the advantages that coupling is simple and the coated beads stable.

Investigation of the ability of several naturally occurring and synthetic polyanions to bind to and potentiate the biological activity of acidic fibroblast growth factor

Publisher: Wiley

Date: 10-1993

DOI: 10.1002/JCP.1041570124

Abstract: The ability of several animal, plant, and bacterial derived polyanions (PAs) as well as synthetic PAs to compete with heparin for the binding of acidic fibroblast growth factor (aFGF) was correlated with their ability to potentiate the mitogenic and neurotrophic actions of this factor. Dextran sulphate, kappa-carrageenan, pentosan sulphate, polyanethole sulfonate, heparin, and fucoidin competed for the heparin binding site on aFGF at relatively low concentrations (< 50 micrograms/ml). lambda-carrageenan, iota-carrageenan, and polyvinyl sulphate exhibited lower affinity for aFGF, whereas hyaluronic acid, dermatan sulphate, chondroitin-6-sulphate, chondroitin-4-sulphate, and uncharged dextran displayed very low or no demonstrable affinity. Potentiation of the mitogenic action of aFGF for Balb/c 3T3 fibroblasts tended to be in general agreement with the aFGF binding affinity of the PAs. However, polyanethole sulfonate, the carrageenans, polyvinyl sulphate, fucoidin, and pentosan sulphate exerted a mitogenic action on the 3T3 cells that was independent of, and in addition to, the ability of these GAGs to potentiate the action of aFGF. The ability to potentiate the neurotrophic action of aFGF for E8 chick ciliary neurons was a general property of those PA with low or no activity in the mitogen assay. Thus hyaluronic acid, dermatan sulphate, chondroitin-4-sulphate, chondroitin-6-sulphate, and even unchanged dextran all potentiated aFGF induced neuronal survival. The differential effects of these PA in potentiating the biological activities of aFGF are discussed in relation to their ability to compete for the heparin-binding site of aFGF.

A new procedure for analysing the relationship between different cell surface antigens.

Publisher: Elsevier BV

Date: 12-1980

DOI: 10.1016/0022-1759(80)90057-5

Abstract: An immunoprecipitation-inhibition procedure is assessing whether different antigenic determinants are carried on the same molecule or on different molecules on cells. The procedure entails (a) exposing NP-40 lysates of cells to antibody against one antigenic specificity (b) removing free antibody and immune complexes by absorption with protein A-bearing S. aureus bacteria (c) adsorption of the NP-40 with a detergent binding resin and (d) measuring the inhibitory activity of the lysates for antibody against another specificity by a rosetting assay. This method has several advantages over the widely used sequential immunoprecipitation procedure.

Cat's whiskers tea (orthosiphon stamineus) extract inhibits growth of colon tumor in nude mice and angiogenesis in endothelial cells via suppressing VEGFR phosphorylation

Publisher: Informa UK Limited

Date: 2012

DOI: 10.1080/01635581.2012.630160

Abstract: Cat's whiskers (Orthosiphon stamineus) is commonly used as Java tea to treat kidney stones including a variety of angiogenesis-dependent diseases such as tumorous edema, rheumatism, diabetic blindness, and obesity. In the present study, antitumor potential of standardized 50% ethanol extract of O. stamineus leaves (EOS) was evaluated against colorectal tumor in athymic mice and antiangiogenic efficacy of EOS was investigated in human umbilical vein endothelial cells (HUVEC). EOS at 100 mg/kg caused 47.62 ± 6.4% suppression in tumor growth, while at 200 mg/kg it caused 83.39 ± 4.1% tumor regression. Tumor histology revealed significant reduction in extent of vascularization. Enzyme-linked immunosorbent assay showed EOS (200 mg/kg) significantly reduced the vascular endothelial growth factor (VEGF) level in vitro (211 ± 0.26 pg/ml cell lysate) as well as in vivo (90.9 ± 2 pg/g tissue homogenate) when compared to the control (378 ± 5 and 135.5 ± 4 pg, respectively). However, EOS was found to be noncytotoxic to colon cancer and endothelial cells. In vitro, EOS significantly inhibited the migration and tube formation of human umbilical vein endothelial cells (HUVECs). EOS suppressed VEGF-induced phosphorylation of VEGF receptor-2 in HUVECs. High performance liquid chromatography (HPLC) analysis of EOS showed high rosmarinic acid contents, whereas phytochemical analysis revealed high protein and phenolic contents. These results demonstrated that the antitumor activity of EOS may be due to its VEGF-targeted antiangiogenicity.

Modification of lymphocyte migration by sulfated polysaccharides

Publisher: Wiley

Date: 1986

DOI: 10.1002/EJI.1830160419

Abstract: The role of sulfated polysaccharides in lymphocyte migration has been analyzed in vivo using lymphocytes labeled with an intracellular DNA-binding fluorochrome Hoechst 33342. The influence of a panel of sulfated polysaccharides on entry (by injecting the sulfated polysaccharide prior to the labeled cells) and displacement from lymphoid organs (by injecting the sulfated polysaccharide after the labeled cells have localized) indicated that different sulfated polysaccharides have selective effects on entry and displacement, and furthermore positioning of subpopulations within organs. Additional experiments suggested that receptors for sulfated polysaccharides on high endothelial venules may interact with complementary structures on lymphocytes. The data supporting this conclusion were: (a) the normal localization behavior of lymphocytes preincubated with sulfated polysaccharides (b) an inverse relationship between the expression of lymphocyte surface receptors for sulfated polysaccharides and the ability of the lymphocytes to enter lymphoid organs and (c) the selective binding of sulfated polysaccharide-coupled fluoresceinated beads to high endothelial venules. In this case only the beads coupled with the sulfated polysaccharides that inhibited entry bound to the high endothelial venules. These findings are discussed in terms of a fundamental cellular recognition system utilizing sulfated polysaccharides.

Chemical and biological properties of bacterial flagellin following iodination and oxidation by chloramine-T

Publisher: Elsevier BV

Date: 09-1972

DOI: 10.1016/0019-2791(72)90160-7

Abstract: Previous studies suggest that hair products containing endocrine disrupting chemicals could alter puberty. We evaluated the association between childhood hair product use and age at menarche in a racially erse study population. We recruited 300 African-American, African-Caribbean, Hispanic, and white women from the New York City metropolitan area who were between 18-77 years of age. Data were collected retrospectively on hair oil, lotion, leave-in conditioner, perm, and other types of hair products used before age 13. Recalled age at menarche ranged from 8 to 19 years. We used multivariable binomial regression to evaluate the association between hair product use and age at menarche (<12 vs. ≥12), adjusting for potential confounders. African-Americans were more likely to use hair products and reached menarche earlier than other racial/ethnic groups. Women reporting childhood hair oil use had a risk ratio of 1.4 (95% confidence interval [CI]: 1.1-1.9) for earlier menarche, adjusting for race/ethnicity and year of birth. Hair perm users had an increased risk for earlier menarche (adjusted risk ratio = 1.4, 95% CI: 1.1-1.8). Other types of hair products assessed in this study were not associated with earlier menarche. Childhood hair oil and perm use were associated with earlier menarche. If replicated, these results suggest that hair product use may be important to measure in evaluating earlier age at menarche.

Anti-self receptors. III. Lack of allelic exclusion and thymic epithelium dependence of H-2L-region-restricted receptors on lymphocytes.

Publisher: Wiley

Date: 06-1981

DOI: 10.1111/J.1365-3083.1981.TB00166.X

Abstract: Murine thymocytes and peripheral lymphocytes bind autologous erythrocytes via H-2L-region-restricted receptors. After inhibiting autorosetting with different erythrocyte sonicates the specificity of these anti-self receptors was examined in F1 hybrid and chimaeric mice. Most F1 lymphocytes simultaneously expressed receptors against both parental haplotypes. Furthermore, analysis of lymphocytes from allogeneic and semi-allogeneic chimaeras clearly demonstrated that radioresistant elements in the recipient thymus did not modify the haplotype specificity of the receptors on donor-derived lymphocytes.

A synthetic heparanase inhibitor reduces proteinuria in passive heymann nephritis

Publisher: Ovid Technologies (Wolters Kluwer Health)

Date: 11-2004

DOI: 10.1097/01.ASN.0000142426.55612.6D

A procedure for removing red cells and dead cells from lymphoid cell suspensions

Publisher: Elsevier BV

Date: 06-1975

DOI: 10.1016/0022-1759(75)90026-5

Abstract: A procedure is described for simultaneously removing red cells and dead cells from lymphoid cell suspensions, based on the observation that when populations of lymphoid cells are centrifuged on a mixture of Isopaque/Ficoll, dead cells and red cells sediment whereas viable cells float. The technique very efficiently removed red cells from a wide range of lymphoid cell suspensions and eliminated lymphocytes killed by mechanical stress, by antibody and complement and by prolonged tissue culture. The depletion of red cells was greater than 99% and the recovery of viable lymphocytes usually greater than 90%, the resulting cell suspensions being around 95-100% viable. The immunological activity of B cells, helper T cells and cytotoxic T cells virtually unimpaired by the separation procedure.

Rapid screening of monoclonal antibodies by spin adherence double immunosorbent test (SADIST)

Publisher: Elsevier BV

Date: 02-1985

DOI: 10.1016/0022-1759(85)90304-7

Abstract: The spin adherence double immunosorbent test (SADIST) is a simple, rapid immunoassay with sensitivity similar to the enzyme-linked immunosorbent assay (ELISA). A 1-step SADIST has been found suitable for rapid screening of hybridomas for antigen-specific monoclonal antibodies (MAb). In this procedure hybridoma supernatants are added to antigen coated microplates followed by commercially available antiglobulin beads. The microplate is immediately centrifuged. Wells containing antigen-specific MAb produce a mat of beads whilst wells without antigen-specific MAb produce a button of beads. No washing or incubation steps are necessary and results are read within minutes of adding beads to test supernatants. By comparison, ELISA tests require several hours to perform with multiple wash steps and further reagent additions. A 2-step SADIST was also assessed. Supernatants are incubated in the microplate as for an ELISA and a wash step precedes the addition of antiglobulin beads. A panel of 117 hybridoma supernatants was selected to assess the suitability of the SADIST techniques for hybridoma screening. The supernatants were added to antigen-coated microplates and SADIST and ELISA tests performed. The SADIST correctly discriminated most hybridoma supernatants that were clearly positive or negative by ELISA. It was also found possible to perform SADIST followed by ELISA tests on the same microplate well without significantly affecting ELISA values.

Dynamic Histone Variant Exchange Accompanies Gene Induction in T Cells

Publisher: Informa UK Limited

Date: 04-2009

DOI: 10.1128/MCB.01590-08

Reactivity of xenogeneic antihuman Ia antisera with the I-C subregion of the murine major histocompatibility complex. I. Genetic studies.

Publisher: Ovid Technologies (Wolters Kluwer Health)

Date: 06-1981

DOI: 10.1097/00007890-198106000-00015

Abstract: Several rabbit xenoantisera raised against human serum Ia antigens have been shown to detect murine lymphocyte antigens. The murine system detected was shown to be polymorphic and expressed on B cells and not T cells. Serological analysis of recombinant H-2 strains demonstrated that two of the antisera recognized products of the I-Cd subregion, and one antiserum recognized products of the I-Cb subregion, thus clearly establishing the existence of the I-C subregion in these two haplotypes. However, as the I-Cd subregion is classically described by the Ia.6 specificity present on T cells and xenoantisera detect a product on B cells, the indication is that the I-C region, like I-A and I-J, complex and contains genes coding for specificities on T cells or B cells.

Reactivity of xenogeneic antihuman Ia antisera with the I-C subregion of the murine major histocompatibility complex. II. Biochemical studies.

Publisher: Ovid Technologies (Wolters Kluwer Health)

Date: 06-1981

DOI: 10.1097/00007890-198106000-00016

Abstract: The biochemical nature of the cross-reactive antigens on murine B cells, detected with xenogeneic antihuman Ia (H.Ia) sera, were examined. These antigens, coded for by the I-C subregion, were found to be carbohydrate in nature and probably exist as glycolipids on the cell surface. Furthermore, sugar inhibition studies demonstrated that the anti-H.Ia sera recognized identical monosaccharide residues on murine and human B cells.

Nonimmune lymphocyte-macrophage interaction

Publisher: Elsevier BV

Date: 05-1985

DOI: 10.1016/0008-8749(85)90009-7

Abstract: This paper describes attempts to determine the molecular basis of the nonimmune interaction between lymphocytes and macrophages. Initial studies revealed that the interaction could be inhibited by simple sugars, six out of the thirty-five tested being inhibitory. Furthermore, the majority of the inhibitory sugars were charged and subsequent studies revealed that some sulfated polysaccharides, notably kappa-carrageenan, were potent inhibitors of the interaction. Further experiments revealed that the lymphocyte-macrophage interaction was indeed mediated by kappa-carrageenan-specific receptors on lymphocytes. The results supporting such a conclusion were as follows: When the interacting cells were preincubated with kappa-carrageenan, it was found that kappa-carrageenan exerted its inhibitory effect at the lymphocyte rather than the macrophage level. Separation of splenocytes into kappa-carrageenan-binding and -nonbinding subpopulations resulted in a corresponding enrichment and depletion of lymphocytes that reacted with macrophages. Lymphocytes were found to express kappa-carrageenan-reactive molecules, these molecules being detected on the surface of lymphocytes by rosetting and in detergent lysates as hemagglutinins. Furthermore, the polyanion specificity of these kappa-carrageenan-specific receptors/hemagglutinins closely resembled the specificity of inhibition of the lymphocyte-macrophage interaction. Pronase-resistant material in macrophage, but not lymphocyte lysates, effectively inhibited both the lymphocyte-macrophage interaction and the recognition of kappa-carrageenan by lymphocytes, suggesting that a kappa-carrageenan-like structure is expressed by macrophages.

Loss of intra-islet heparan sulfate is a highly sensitive marker of type 1 diabetes progression in humans

Publisher: Public Library of Science (PLoS)

Date: 07-02-2018

DOI: 10.1371/JOURNAL.PONE.0191360

Detection of developmentally controlled plasma membrane antigens of Dictyostelium discoideum cells in SDS-polyacrylamide gels.

Publisher: Wiley

Date: 15-11-1978

DOI: 10.1016/0014-5793(78)81031-X

Interplay between Chromatin Remodeling and Epigenetic Changes during Lineage-Specific Commitment to Granzyme B Expression

Publisher: The American Association of Immunologists

Date: 12-2009

DOI: 10.4049/JIMMUNOL.0901522

Abstract: The role of chromatin remodeling and histone posttranslational modifications and how they are integrated to control gene expression during the acquisition of cell-specific functions is poorly understood. We show here that following in vitro activation of CD4+ and CD8+ T lymphocytes, both cell types show rapid histone H3 loss at the granzyme B (gzmB) proximal promoter region. However, despite the gzmB proximal promoter being remodeled in both T cell subsets, only CD8+ T cells express high levels of gzmB and display a distinct pattern of key epigenetic marks, notably differential H3 acetylation and methylation. These data suggest that for high levels of transcription to occur a distinct set of histone modifications needs to be established in addition to histone loss at the proximal promoter of gzmB.

Mice deficient in the putative phospholipid flippase Atp11c exhibit altered erythrocyte shape, anemia, and reduced erythrocyte life span

Publisher: Elsevier BV

Date: 07-2014

DOI: 10.1074/JBC.C114.570267

An improved colorimetric assay for T cell cytotoxicity in vitro

Publisher: Elsevier BV

Date: 03-1984

DOI: 10.1016/0022-1759(84)90151-0

Abstract: An improved colorimetric assay for estimation of cytotoxic T (Tc) cells is described. The method involves staining thioglycollate-induced macrophage targets with the dye neutral red prior to addition of cytotoxic T cells and estimating macrophage survival at the end of the assay by measuring dye remaining in viable targets. The method using macrophage targets is more sensitive than the 51Cr release assay employing macrophages or a variety of other targets. It may be used to detect alloreactive and H-2 restricted Tc cells in both short-term (4 h) and long-term (24 h) assays and overcomes some variability encountered with a previously described colorimetric procedure. Furthermore, the method is cheap, fast, reliable and avoids the use of radioactivity.

Adenovirus-induced alterations of the cell growth cycle: a requirement for expression of E1A but not of E1B

Publisher: American Society for Microbiology

Date: 1983

DOI: 10.1128/JVI.45.1.192-199.1983

Abstract: Mutants dl312, dl314, hr1, and hr3 with mutations in region E1A of adenovirus type 5 were defective for the induction of cell cycle abnormalities detectable by flow cytometry, cell DNA replication, thymidine kinase production, and chromosome aberrations and did not synthesize the viral DNA-binding protein (E2A) in rat cells. dl311, a leaky E1A mutant, induced cell cycle effects at high multiplicity in only one of three experiments, and synthesized the DNA-binding protein. hr7 (E1B) gave a wild-type response in all tests. dl313 was also positive in all tests, although it induced fewer polyploid cells than did wild-type virus, probably because of the leftward extension of the dl313 E1B deletion into E1A. sub315 and sub316, with mutations which also span the E1A-E1B border, synthesized DNA-binding protein, but caused no cell cycle alterations detectable by flow cytometry in rat or mouse cells. Although the participation of other viral early regions cannot be completely excluded, our results suggest that alteration of cell cycle progression is a direct effect of E1A unrelated to its control of other viral early regions, and may be the function of E1A in transformation.

The Relationship Between Humoral and Cell-Mediated Immunity

Publisher: Wiley

Date: 12-1972

DOI: 10.1111/J.1600-065X.1972.TB00059.X

Platelets and P-selectin control tumor cell metastasis in an organ-specific manner and independently of NK cells

Publisher: American Association for Cancer Research (AACR)

Date: 15-09-2012

DOI: 10.1158/0008-5472.CAN-11-4010

Abstract: The prometastatic role of platelets has long been recognized with proposed mechanisms of action including shielding tumor cells from natural killer (NK) cell destruction and aiding endothelial attachment and extravasation of tumor cells with platelet P-selectin being implicated in these processes. However, many aspects of the prometastatic function of platelets remain unclear. In this study, we used mouse models of metastatic breast cancer and melanoma to investigate the platelet effect, focusing on organ specificity, the relationship with NK cells and the relative importance of platelet-derived versus endothelial-derived P-selectin. We found that platelets promote lung metastasis in the absence of NK cells in both acute and spontaneous metastasis models. In addition, the prometastatic action of platelets was found to be organ specific, clearly enhancing lung metastasis but not affecting B16F1 liver metastasis, in fact, liver metastasis was enhanced in the absence of platelets. Furthermore, the profound antimetastatic activity of NK cells was equally effective in the presence or absence of platelets and chronologically distinct from the prometastatic role of platelets. Finally, it was shown that endothelial-derived P-selectin is just as important as platelet-derived P-selectin in promoting lung metastasis and also plays an important role in liver metastasis. Taken together, our findings help clarify the roles of platelets, NK cells and P-selectin in metastasis, and they identify P-selectin as an attractive therapeutic target for preventing metastasis in multiple organs. Cancer Res 72(18) 4662–71. ©2012 AACR.

The Transcriptional Repressor Bcl-6 Directs T Follicular Helper Cell Lineage Commitment

Publisher: Elsevier BV

Date: 09-2009

DOI: 10.1016/J.IMMUNI.2009.07.002

Abstract: Follicular helper T (Tfh) cells provide selection signals to germinal center B cells, which is essential for long-lived antibody responses. High CXCR5 and low CCR7 expression facilitates their homing to B cell follicles and distinguishes them from T helper 1 (Th1), Th2, and Th17 cells. Here, we showed that Bcl-6 directs Tfh cell differentiation: Bcl-6-deficient T cells failed to develop into Tfh cells and could not sustain germinal center responses, whereas forced expression of Bcl-6 in CD4(+) T cells promoted expression of the hallmark Tfh cell molecules CXCR5, CXCR4, and PD-1. Bcl-6 bound to the promoters of the Th1 and Th17 cell transcriptional regulators T-bet and RORgammat and repressed IFN-gamma and IL-17 production. Bcl-6 also repressed expression of many microRNAs (miRNAs) predicted to control the Tfh cell signature, including miR-17-92, which repressed CXCR5 expression. Thus, Bcl-6 positively directs Tfh cell differentiation, through combined repression of miRNAs and transcription factors.

Inhibition of in vitro antibody synthesis by cyclophosphamide-induced suppressor cells.

Publisher: Elsevier BV

Date: 04-1980

DOI: 10.1016/0008-8749(80)90232-4

Abstract: Dengue infections have become a huge threat to public health systems in developing countries. Data on seroprevalence and incidence of dengue infections are lacking from rural regions of India. The objective of present study was to investigate the seroprevalence and incidence of dengue infection utilizing repeated serosurveys from a rural region of Maharashtra, Western India. In the present study, 819 children between ages 5 to 15 years from 21 villages in Pune District of Maharashtra, India were s led in 2014 and 2016. The sera were tested for the presence of dengue specific IgG using an indirect IgG ELISA kit. Overall seroprevalence of dengue was 15.3% (95% confidence intervals (CI) 12.9-17.8%) in 2014 and 20.5% (95% CI 17.8-23.4%) in 2016. Among the 694 children who were seronegative at baseline (2014), 78 seroconverted. Overall incidence rate of primary dengue was 54.2 infections/1000 children years (95% CI 43.0-67.3). Incidence of primary dengue infection was higher in children from urbanized villages compared to rural villages (Incidence rate ratio (IRR) 2.6 (95% CI 1.3-5.2)). In rural villages, incidence of primary dengue infection was higher in children aged 10 years or above as compared to those aged below 10 years (IRR 9.75 (95% CI 1.21-77.9). The study provides the incidence rates of primary dengue infections from a rural region of India. More multi centric studies investigating the incidence of dengue will provide accurate estimate of incidence of dengue and help formulate well directed policies. The results also suggest that urbanization and transitions in demographic settings might favour dengue outbreaks in rural regions and these regions need to be targeted for vector control measures.

Anti-self receptors. I. Direct detection of H-2L region-restricted receptors on murine thymocytes.

Publisher: Rockefeller University Press

Date: 03-1980

DOI: 10.1084/JEM.151.3.553

Abstract: A high proportion (20--50%) of murine thymocytes form rosettes with either syngeneic or allogeneic erythrocytes. The specificity of this interaction was investigated by measuring the ability of different erythrocyte sonicates to inhibit rosette formation. With erythrocyte sonicates from recombinant mouse strains it was demonstrated that rosetting with syngeneic erythrocytes was mediated by H-2L and/or H-2D region-restricted receptors. The specificity of autorosetting was directly mapped to the H-2L region by the inability of erythrocyte sonicates from the BALB/c-H-2dm2 mutant, an H-2L-deletion mutant, to inhibit the rosetting of wild-type (BALB/c) thymocytes. The B10,2D2-H-2dm1 mutant, which has substantially modified H-2L and H-2D antigens, supported this conclusion. Furthermore, anti-H-2L sera were able to specifically block the inhibition of rosetting by erythrocyte sonicates. The above procedures clearly implicated the H-2L region in the thymocyte rosetting of d and k haplotypes. With the s haplotype the rosetting receptor was mapped to the H-2L/H-2D region, whereas with the b and q haplotypes rosetting was only mapped to the D end of the H-2 complex. This study also suggested complete cross-reaction between the thymocyte receptors carried by the k and d haplotypes, whereas the receptors of b, q, and s haplotypes were haplotype specific. In addition, the inhibition assay indicated that the rosetting of thymocytes with allogeneic and xenogeneic (rat) erythrocytes was mediated by a receptor primarily directed against self-H-2L. Finally, the critical role played by the H-2L region in this rosetting phenomenon was demonstrated by the inability of thymocytes from the H-2L-deletion mutant (H-2dm2) to rosette with syngeneic, allogeneic (rat) erythrocytes.

Heparanase in primary human osteoblasts

Publisher: Wiley

Date: 05-08-2010

DOI: 10.1002/JOR.21138

Abstract: Heparanase (HPSE) is known to be involved in fracture repair in mice, but its presence and function in human bone formation remains unclear. Our aim was to determine the expression of HPSE in human bone forming osteoblasts and to better understand its role in osteogenesis. HPSE protein expression and enzymatic activity were demonstrated in osteoblasts isolated from trabecular bone specimens of patients with osteoporosis (OP) and from healthy subjects, although the levels differed markedly. Thus, low levels of HPSE expression were observed in osteoporotic osteoblasts, including in the nucleus compared to those from healthy subjects. Notably, HPSE gene expression was associated with alkaline phosphatase (ALP) activity, the bone turnover marker. Gene profile studies demonstrated that osteogenic genes were downregulated in osteoporotic osteoblasts. We further exposed osteoblasts to exogenous HPSE and found that the level of histone H3 phosphorylation was increased. We provide evidence, for the first time, demonstrating that HPSE expresses and functions in human osteoblasts. Our data suggest that previously undescribed function of HPSE-mediated osteoblastogenesis through regulation of osteogenic gene expression and histone H3 modification. HPSE upregulation may be a novel therapeutic approach in the prevention and treatment of OP.

Extracellular histones are a target in myocardial ischaemia–reperfusion injury

Publisher: Oxford University Press (OUP)

Date: 20-04-2021

DOI: 10.1093/CVR/CVAB139

Abstract: Acute myocardial infarction causes lethal cardiomyocyte injury during ischaemia and reperfusion (I/R). Histones have been described as important Danger Associated Molecular Proteins (DAMPs) in sepsis. The objective of this study was to establish whether extracellular histone release contributes to myocardial infarction. Isolated, perfused rat hearts were subject to I/R. Nucleosomes and histone-H4 release was detected early during reperfusion. Sodium-β-O-Methyl cellobioside sulfate (mCBS), a newly developed histone-neutralizing compound, significantly reduced infarct size whilst also reducing the detectable levels of histones. Histones were directly toxic to primary adult rat cardiomyocytes in vitro. This was prevented by mCBS or HIPe, a recently described, histone-H4 neutralizing peptide, but not by an inhibitor of TLR4, a receptor previously reported to be involved in DAMP-mediated cytotoxicity. Furthermore, TLR4-reporter HEK293 cells revealed that cytotoxicity of histone H4 was independent of TLR4 and NF-κB. In an in vivo rat model of I/R, HIPe significantly reduced infarct size. Histones released from the myocardium are cytotoxic to cardiomyocytes, via a TLR4-independent mechanism. The targeting of extracellular histones provides a novel opportunity to limit cardiomyocyte death during I/R injury of the myocardium.

Sendai virus stimulates chemiluminescence in mouse T and B lymphocytes

Publisher: Wiley

Date: 06-1980

DOI: 10.1002/EJI.1830100613

EFFECTS OF THYMUS-INDEPENDENT (B) CELLS AND THE H-2 GENE COMPLEX ON ANTIVIRAL FUNCTION OF IMMUNE THYMUS-DERIVED (T) CELLS

Publisher: Wiley

Date: 06-1975

DOI: 10.1038/ICB.1975.19

Abstract: Antiviral activity in vivo exerted by ectromelia virus-immune spleen cells transferred to ectromelia-infected recipients and cytotoxicity against virus-infected target cells in vitro were both properties of non-immunoglobulin (Ig)-bearing cells (which included T cells). Ig-bearing cells, including thymus-independent (B) cells and antibody-secreting cells, were much less active in vivo when injected alone and tended to block rather than lify the effect triggered by T cells. Ig-bearing cells were also slightly active in vitro, possibly because some T cells have detectable Ig. Antiviral effects in cell transfer experiments were seen only when immune cell donors and infected recipients shared the same H-2 gene complex. These results are consistent with the hypothesis that the T cell response to ectromelia infection is directed against specific virus-induced change(s) in antigen(s), specified by gene(s) in the H-2 complex, which appear in virus-infected cells.

Neuropilin 1-Sema Signaling Regulates Crossing of Cingulate Pioneering Axons during Development of the Corpus Callosum

Publisher: Oxford University Press (OUP)

Date: 08-04-2009

DOI: 10.1093/CERCOR/BHP027

Simple model for self-non-self-discrimination in invertebrates

Publisher: Springer Science and Business Media LLC

Date: 06-1977

DOI: 10.1038/267711A0

Histidine-rich glycoprotein is a novel plasma pattern recognition molecule that recruits IgG to facilitate necrotic cell clearance via FcγRI on phagocytes

Publisher: American Society of Hematology

Date: 25-03-2010

DOI: 10.1182/BLOOD-2009-07-234013

Abstract: Under normal physiologic conditions, necrotic cells resulting from tissue injury are rapidly removed from the circulation and tissues by phagocytes, thus preventing the exposure of intracellular antigenic and immunostimulatory molecules that can aid the development of autoimmune disease. Histidine-rich glycoprotein (HRG), a relatively abundant plasma glycoprotein, has a multidomain structure that can interact with many ligands including components of the fibrinolytic and immune systems. Recently, it has been reported that HRG can bind strongly to cytoplasmic ligand(s) exposed in necrotic cells to enhance clearance by phagocytes. Here we describe the molecular mechanisms underpinning this process. A complex consisting of both HRG and immunoglobulin G (IgG) was found as necessary to aid necrotic cell uptake by monocytes, predominantly via an FcγRI-dependent mechanism. The findings in this study also show that HRG can potentially interact with anionic phospholipids exposed in necrotic cells. Furthermore, the enhanced phagocytosis of necrotic cells induced by HRG-IgG complexes triggers phagocytes to release proinflammatory cytokines such as interleukin-8 and tumor necrosis factor. Thus, HRG has the unique property of complexing with IgG and facilitating a proinflammatory innate immune response to promote the clearance of necrotic cells.

The low molecular weight heparan sulfate-mimetic, PI-88, inhibits cell-to-cell spread of herpes simplex virus

Publisher: Elsevier BV

Date: 07-2004

DOI: 10.1016/J.ANTIVIRAL.2004.01.001

A detailed serological analysis of a xenogeneic anti-Ia serum

Publisher: Springer Science and Business Media LLC

Date: 12-1978

DOI: 10.1007/BF01563908

Histidine-rich glycoprotein binds to cell-surface heparan sulfate via its N-terminal domain following Zn²⁺ chelation

Publisher: Elsevier BV

Date: 07-2004

DOI: 10.1074/JBC.M401996200

Heparanase: A key enzyme involved in cell invasion

Publisher: Elsevier BV

Date: 03-2001

DOI: 10.1016/S0304-419X(01)00017-8

Targeting Dendritic Cells with Antigen-Containing Liposomes

Publisher: American Association for Cancer Research (AACR)

Date: 15-06-2004

DOI: 10.1158/0008-5472.CAN-04-0138

Abstract: Dendritic cells (DCs) are potent stimulators of immunity, and DCs pulsed with tumor antigen ex vivo have applications in tumor immunotherapy. However, DCs are a small population of cells, and their isolation and pulsing with antigen can be impractical. Here we show that a crude preparation of plasma membrane vesicles (PMV) from the highly metastatic murine melanoma (B16-OVA) and a surrogate tumor antigen (OVA) can be targeted directly to DCs in vivo to elicit functional effects. A novel metal-chelating lipid, 3(nitrilotriacetic acid)-ditetradecylamine, was incorporated into B16-OVA-derived PMV, allowing recombinant hexahistidine-tagged forms of single chain antibody fragments to the DC surface molecules CD11c and DEC-205, to be conveniently “engrafted” onto the vesicle surface by metal-chelating linkage. The modified PMV, or similarly engrafted synthetic stealth liposomes containing OVA or OVA peptide antigen, were found to target DCs in vitro and in vivo, in experiments using flow cytometry and fluorescence confocal microscopy. When used as vaccines in syngeneic mice, the preparations stimulated strong B16-OVA-specific CTL responses in splenic T cells and a marked protection against tumor growth. Protection was dependent on the simultaneous delivery of both antigen and a DC maturation or “danger signal” signal (IFN-γ or lipopolysaccharide). Administration of the DC-targeting vaccine to mice challenged with B16-OVA cells induced a dramatic immunotherapeutic effect and prolonged disease-free survival. The results show that the targeting of antigen to DCs in this way is highly effective at inducing immunity and protection against the tumor, with protection being at least partially dependent on the eosinophil chemokine eotaxin.

The lymphocyte surface. II. Separation of Fc receptor, C'3 receptor and surface immunoglobulin-bearing lymphocytes

Publisher: The Royal Society

Date: 27-08-1974

DOI: 10.1098/RSPB.1974.0061

Abstract: A one-step separation procedure is described for both depleting and obtaining in pure form Fc receptor (FcRL), C'3 receptor (CRL) and surface immunoglobulin bearing (IgL) lymphocytes from rat lymphoid populations. The method is a modification of the Bӧyum (1968) technique for separating lymphocytes from whole blood by sedimentation on Ficoll/Isopaque, and is based on the fact that when a lymphocyte forms a rosette with sensitized erythrocytes it will sediment with the red cells rather than float with the non-rosetting lymphocytes. The technique is 99.5% efficient at depleting thoracic duct lymphocytes (TDL) of FcRL, CRL and IgL and these subpopulations can be recovered 93-98% pure. The total recovery of lymphocytes applied is usually 90% and the separated lymphocytes are 95% viable. This technique allowed the cellular distribution of Fc receptors, C'3 receptors and surface Ig to be determined. It was found that ( a ) Almost all CRL carry surface Ig, although a very small sub-population of CRL (0.2-0.8%) which lacks surface Ig could regularly be detected. ( b ) A substantial proportion of IgL (12-25%) lacks C'3 receptors. ( c ) IgL and CRL which lack Fc receptors are more frequent in spleen and lymph nodes than in TDL. The proportion of this subpopulation increases in TDL after prolonged thoracic duct drainage. ( d ) Some FcRL exist which lack both C'3 receptors and surface Ig. These cells are more evident in TDL after prolonged thoracic duct drainage and in lymph nodes (20-30% of FcRL) than in early TDL or spleen (5-10% of FcRL). ( e ) The thymus contains very few FcRL, CRL or IgL. ( f ) A large population of lymphocytes exists in B rats (32-42% of TDL) which is killed by an anti-B serum but which lacks surface Ig. These cells are much less frequent in normal TDL ( 5%) and probably also lack Fc and C'3 receptors. ( g ) Large lymphocytes probably shed their Fc and C'3 receptors, but retain their surface Ig, during S-phase. ( h ) Studies on a secondary anti-DNP response showed that a substantial proportion of direct and indirect plaque forming cells (PFC) in the spleen express Fc receptors, whereas only indirect PFC carry C'3 receptors. Virtually all PFC ( 98%) possess surface Ig.

Histidine-rich glycoprotein specifically binds to necrotic cells via its amino-terminal domain and facilitates necrotic cell phagocytosis

Publisher: Elsevier BV

Date: 10-2005

DOI: 10.1074/JBC.M504384200

The lymphocyte surface. I. Relation between Fc receptors, C'3 receptors and surface immunoglobulin

Publisher: The Royal Society

Date: 27-08-1974

DOI: 10.1098/RSPB.1974.0060

Abstract: Fc receptors, C'3 receptors and immunoglobulin (Ig) were detected on the surface of rat thoracic duct lymphocytes by a series of rosetting procedures. This paper describes the rosetting methods and some of the general properties of these cell surface components. It was found that the Fc receptors were blocked by antigen-antibody complexes and anti─Ag-B antibodies, they were lost in vitro at 37 °C, and they were not detected in the presence of 10 -4 M azide. In contrast, the C'3 receptors were destroyed by treatment with trypsin, were insensitive to azide, and were not blocked by antigen-antibody complexes or anti─Ag-B anti-bodies. Both receptors were detected readily at 20 or 37 °C but not at 0 °C. Neither the Fc or C'3 receptors capped with surface Ig. From these differences in behaviour it was concluded that the Fc receptors, C'3 receptors and surface Ig probably represent separate molecules on the lymphocyte surface.

A rapid, automated colorimetric assay for measuring antibody binding to cell surface antigens

Publisher: Elsevier BV

Date: 11-1983

DOI: 10.1016/0022-1759(83)90433-7

Abstract: An automated, colorimetric procedure is described for detecting antibodies specific for cell surface antigens. The procedure entails (a) coating the wells of 96-well microplates with either protein A or anti-immunoglobulin antibodies and (b) preincubating either the microplate or target cells with the test antibody. Target cells which react with the test antibody bind to the wells of the microplate and bound cells are quantitated by staining with the dye Rose Bengal. A microplate spectrophotometer is used to measure absorbance in each well of the plate, providing a rapid, automated measure of antibody titre. The assay is simple to perform, uses readily available reagents and gives comparable sensitivity to rosetting assays. With these features, and the capacity for handling large numbers of trays quickly, this method has obvious advantages in screening for antibody activity in culture supernatants of hybridoma clones.

C8c-C15 monoseco-analogues of the phenanthroquinolizidine alkaloids julandine and cryptopleurine exhibiting potent anti-angiogenic properties

Publisher: Elsevier BV

Date: 2006

DOI: 10.1016/J.BMCL.2005.09.032

Abstract: Four enantiomerically pure monoseco-analogues, 5, 7, 9, and 11, of the phenanthroquinolizidine alkaloid julandine (1) and four of congener cryptopleurine (2), viz. compounds 6, 8, 10, and 12, have been prepared and subjected to preliminary biological evaluation. These analogues show dramatically reduced cytotoxicity compared with the parent system 2 but they are, nevertheless, potent anti-angiogenic agents.

Fluorescent target array T helper assay: A multiplex flow cytometry assay to measure antigen-specific CD4⁺ T cell-mediated B cell help in vivo

Publisher: Elsevier BV

Date: 2013

DOI: 10.1016/J.JIM.2012.10.013

Abstract: CD4(+) T cells play a central role in regulating the immune response. Their effector function is commonly assessed by their capacity to secrete cytokines detected by ELISPOT and intracellular cytokine staining. However, one aspect of their effector function that is often overlooked is their ability to help activation of cognate B cells directly, a process that is initiated through the engagement of their T cell-receptor (TCR) with cognate peptide presented on major histocompatibility complex class II (MHC-II) molecules by B cells. Here we report a method to monitor CD4(+) T cell-mediated B cell help in vivo using a multiplex high throughput assay. This assay utilizes a fluorescent target array (FTA), which is composed of lymphocytes labeled with numerous (>200) unique fluorescence signatures that can be delineated in a single recipient animal based on combination labeling with the three vital dyes carboxyfluorescein diacetate succinimidyl ester (CFSE), CellTrace Violet (CTV) and Cell Proliferation Dye eFluor 670 (CPD). By pulsing different B cell populations in a FTA with titrated amounts of cognate MHC-II binding peptides, CD4(+) T cell help could be assessed by measuring induction of the B cell activation markers CD69 and CD44 by antibody labeling and flow cytometry. We call this the "FTA T helper assay", and have found it to be a robust and sensitive assay to measure CD4(+) T cell helper activity across a multitude of peptide-pulsed B "target" cells in real time in vivo. Furthermore, the technique can be used simultaneously with the FTA killing assay that measures cytotoxic T cell function, to provide a comprehensive tool for measuring both CD4(+) and CD8(+) T cell activity during an immune response in vivo.

Anti-HIV-1 Activity of Chemically Modified Heparins: Correlation between Binding to the V3 Loop of gp120 and Inhibition of Cellular HIV-1 Infection in Vitro

Publisher: American Chemical Society (ACS)

Date: 07-06-1994

DOI: 10.1021/BI00188A029

Abstract: Chemically modified heparins were tested for their activities in (i) inhibiting HIV-1 replication in vitro and (ii) inhibiting the binding to recombinant HIV-1 gp120 of monoclonal antibodies specific for the V3 loop. The results reveal that N-desulfation reduces activity, although this is largely restored on N-acetylation. Selective O-desulfation also markedly reduces activity, whereas carboxyl reduction has little effect. Overall these results show that the anti-HIV-1 activity of heparin does not depend simply on negative density, and indicate instead that particular structures, notably O-sulfates, are involved. Our studies reveal that for chemically modified heparins and heparin-derived fragments there is a striking correlation between anti-HIV-1 activity in vitro and binding to the V3 loop of gp120 in solid phase ELISA. This strongly suggests that the heparin exerts its anti-HIV-1 activity by binding to the V3 loop of gp120.

A sensitive and quantitative fluorescence assay for cell surface antigens.

Publisher: Elsevier BV

Date: 12-1981

DOI: 10.1016/0022-1759(81)90283-0

Abstract: A procedure is described for the assay of cell surface antigens based on quantitative fluorometry. Fluorescent immunospheres are coupled with sheep anti-mouse immunoglobulins or Protein A and used to detect specific antibody bound to target cells. The fluorescent sphere assay described here offers 16--128-fold greater sensitivity than complement mediated lysis or Protein A radioimmune assays and comparable sensitivity to rosetting assays. In addition, the assay is simple to perform, uses commercially available reagents and is completely objective in that a common laboratory fluorometer is used to obtain fluorescence measurements.

Regulation of the immune response by antibody. III. The effect of macrophage cytophilic antibody on humoral and cell-mediated immunity

Publisher: Elsevier BV

Date: 12-1972

DOI: 10.1016/0008-8749(72)90102-5

Abstract: Ineffective parenting practices may maintain or exacerbate attention deficit/hyperactivity disorder (ADHD) symptoms and shape subsequent development of disruptive behavior disorders (DBD's) in youth with ADHD. Recent theoretical models have suggested that parenting may exert effects on ADHD via its role in child temperament. The current study aimed to evaluate the indirect effects of parenting dimensions on child ADHD symptoms via child temperament. Youth ages 6-17 years (N = 498 50.4 % ADHD, 55 % male) completed a multi-stage, multi-informant assessment that included parent, child, and teacher report measures of parenting practices, child temperament, and ADHD symptoms. Statistical models examined the direct and indirect effects of maternal and paternal involvement, poor supervision, and inconsistent discipline on inattention and hyperactivity-impulsivity via child temperament and personality traits. Results indicated differential patterns of effect for negative and positive parenting dimensions. First, inconsistent discipline exerted indirect effects on both ADHD symptom dimensions via child conscientiousness, such that higher levels of inconsistency predicted lower levels of conscientiousness, which in turn, predicted greater ADHD symptomatology. Similarly, poor supervision also exerted indirect effects on inattention via child conscientiousness as well as significant indirect effects on hyperactivity-impulsivity via its impact on both child reactive control and conscientiousness. In contrast, primarily direct effects of positive parenting (i.e., involvement) on ADHD emerged. Secondary checks revealed that similar pathways may also emerge for comorbid disruptive behavior disorders. Current findings extend upon past work by examining how parenting practices influence child ADHD via with-in child mechanisms and provide support for multi-pathway models accounting for heterogeneity in the disorder.

A multimeric form of soluble recombinant sheep LFA-3 (CD58) inhibits human T-cell proliferation

Publisher: Wiley

Date: 09-1997

DOI: 10.1046/J.1365-2567.1997.00317.X

Abstract: The rosetting of T cells by sheep erythrocytes is mediated through the interaction of the CD2 molecule on T cells with T11TS, a molecule on sheep erythrocytes homologous to lymphocyte function-associated antigen-3 (LFA-3, CD58). We cloned a T11TS cDNA from sheep leucocyte mRNA which encodes a soluble molecule comprising the distal D1 and the D2 extracellular domains, but not the transmembrane domain. cDNA for this soluble D1 + D2 form of sheep LFA-3 (sLFA-3) was expressed in Escherichia coli and the properties of the purified recombinant protein were assessed by inhibition of T-cell rosette formation. sLFA-3 inhibited rosette formation, but its activity was low, 50% inhibition occurring at 25 micrograms/ml, consistent with the observed low binding avidity of fluorescein isothiocyanate (FITC)-labelled sLFA-3, sLFA-3 was made multimeric to increase its affinity, by crosslinking biotinylated sLFA-3 to streptavidin-biotinylated dextran complexes. The binding of crosslinked sLFA-3 multimers, tested by fluorescence-activated cell sorting (FACS) analysis, was significantly increased compared to sLFA-3 monomers. Competition with monoclonal antibodies demonstrated that multimeric sLFA-3 bound to the T11(1) epitope on CD2. The multimeric form of sLFA-3 was significantly more potent than the monomer in inhibiting proliferation of human T cells in response to purified protein derivative (PPD), tetanus toxoid (TT) or allogeneic cells. Multimeric sLFA-3 might, therefore, have potential as an immunotherapeutic agent to inhibit and/or anergize antigen-specific T-cell responses.

Surface properties of cells involved in antibody-dependent cytotoxicity

Publisher: Elsevier BV

Date: 02-1976

DOI: 10.1016/0008-8749(76)90051-4

Calcein: A novel marker for lymphocytes which enter lymph nodes

Publisher: Wiley

Date: 1992

DOI: 10.1002/CYTO.990130710

Abstract: Previous studies have identified unique cell surface antigens which are associated with the specific binding of lymphocytes to high endothelial venules (HEV). Evidence is presented in this paper which demonstrates that uptake of the fluorescent dye calcein by lymphocytes represents an additional marker for the lymph node homing subpopulation of lymphocytes. Calcein exhibits a characteristic ability to label lymphocytes differentially into two distinct populations, based on fluorescence intensity, that does not occur with three other structurally related, fluorescein-based dyes. In vivo lymphocyte migration studies revealed that cells displaying the "dull" fluorescence phenotype, although entering all lymphoid organs examined, preferentially homed to the lymph nodes, particularly the popliteal lymph node (PLN). By contrast, lymphocytes displaying the "bright" phenotype were essentially excluded from entering lymphoid organs, where entry is HEV dependent, but were observed entering spleen, where entry is HEV independent. Furthermore, a high proportion (76.5%) of lymphocytes displaying the dull fluorescence phenotype expressed the PLN homing receptor MEL-14. Based on these observations it is suggested that calcein uptake may be a marker for general membrane properties, such as fluidity and plasticity, essential for the passage of lymphocytes through HEV.

Cleavage of bacterial flagellin with cyanogen bromide. Antigenic properties of the protein fragments

Publisher: Portland Press Ltd.

Date: 07-1969

DOI: 10.1042/BJ1130501

Abstract: 1. Four polypeptide fragments, obtained by cyanogen bromide treatment of the protein flagellin from Salmonella adelaide, were tested for their antigenic activity by using them as inhibitors in three different assays: bacterial immobilization, haemagglutination of sensitized erythrocytes and quantitative micro precipitation. Immunodiffusion studies were also performed on the protein fragments. 2. Cleavage of the flagellin molecule in this way gave no detectable loss of antigenic determinants. Fragment A (mol.wt. 18000), the largest of the polypeptides, contained all the antigenic specificities present on flagellin that were recognized by the antisera used. In one test, fragment B (mol.wt. 12000) also contained antigenic activity to an extent not easily explainable by contamination with fragment A. Fragments C (mol.wt. 5500) and D (mol.wt. 4500) appeared to be antigenically inactive.

The Structural Proteins of Chick Embryo Lethal Orphan Virus (Fowl Adenovirus Type 1)

Publisher: Microbiology Society

Date: 10-1984

DOI: 10.1099/0022-1317-65-10-1803

Abstract: Chick embryo lethal orphan (CELO) virus (fowl adenovirus type 1) contains at least 14 structural proteins with polypeptide molecular weights ranging from 100K to about 6K. A nomenclature of the CELO virion polypeptides is presented and the molar proportion of each polypeptide has been estimated. The CELO virus pentons were specifically released from the virion by dialysis against borate-based calcium-magnesium saline. The penton base (polypeptide III, mol. wt. 92K) and two fibres were separated, characterized and their polypeptides were correlated with their morphological positions in the virion. Peptide mapping suggested that the long fibre (polypeptide IV, mol. wt. 65K), and the short fibre (polypeptide VII, mol. wt. 44.5K) were not related in their primary sequences and are therefore probably encoded by separate genes. The time course of synthesis of the CELO virion polypeptides indicated that, like their mammalian adenovirus counterparts, they are synthesized late (after viral DNA replication).

Molecular mechanisms of late apoptotic/necrotic cell clearance

Publisher: Springer Science and Business Media LLC

Date: 18-12-2010

DOI: 10.1038/CDD.2009.195

Abstract: Phagocytosis serves as one of the key processes involved in development, maintenance of tissue homeostasis, as well as in eliminating pathogens from an organism. Under normal physiological conditions, dying cells (e.g., apoptotic and necrotic cells) and pathogens (e.g., bacteria and fungi) are rapidly detected and removed by professional phagocytes such as macrophages and dendritic cells (DCs). In most cases, specific receptors and opsonins are used by phagocytes to recognize and bind their target cells, which can trigger the intracellular signalling events required for phagocytosis. Depending on the type of target cell, phagocytes may also release both immunomodulatory molecules and growth factors to orchestrate a subsequent immune response and wound healing process. In recent years, evidence is growing that opsonins and receptors involved in the removal of pathogens can also aid the disposal of dying cells at all stages of cell death, in particular plasma membrane-damaged cells such as late apoptotic and necrotic cells. This review provides an overview of the molecular mechanisms and the immunological outcomes of late apoptotic/necrotic cell removal and highlights the striking similarities between late apoptotic/necrotic cell and pathogen clearance.

Heparan sulfate as a receptor for poxvirus infections and as a target for antiviral agents

Publisher: Microbiology Society

Date: 10-2017

DOI: 10.1099/JGV.0.000921

Abstract: To establish the importance of virus-heparan sulfate (HS) interactions in virus infectivity, the poxvirus vaccinia virus (VACV) was used, as it binds HS and has both enveloped virus (EV) and non-enveloped mature virus (MV) forms. Initial studies showed that heparin inhibited plaque formation by both MV-rich WR and EV-rich IHD-J strains of VACV, with the EV-rich strain also losing trademark 'comet'-shaped plaques. However, using GFP-tagged EV and MV forms of VACV, based on IC

Chromatin-Associated Protein Kinase C-θ Regulates an Inducible Gene Expression Program and MicroRNAs in Human T Lymphocytes

Publisher: Elsevier BV

Date: 03-2011

DOI: 10.1016/J.MOLCEL.2011.02.030

Abstract: Studies in yeast demonstrate that signaling kinases have a surprisingly active role in the nucleus, where they tether to chromatin and modulate gene expression programs. Despite these seminal studies, the nuclear mechanism of how signaling kinases control transcription of mammalian genes is in its infancy. Here, we provide evidence for a hitherto unknown function of protein kinase C-theta (PKC-θ), which physically associates with the regulatory regions of inducible immune response genes in human T cells. Chromatin-anchored PKC-θ forms an active nuclear complex by interacting with RNA polymerase II, the histone kinase MSK-1, and the adaptor molecule 14-3-3ζ. ChIP-on-chip reveals that PKC-θ binds to promoters and transcribed regions of genes, as well as to microRNA promoters that are crucial for cytokine regulation. Our results provide a molecular explanation for the role of PKC-θ not only in normal T cell function, but also in circumstances of its ectopic expression in cancer.

The role of heparan sulphate in inflammation

Publisher: Springer Science and Business Media LLC

Date: 18-08-2006

DOI: 10.1038/NRI1918

Abstract: The polysaccharide heparan sulphate is ubiquitously expressed as a proteoglycan in extracellular matrices and on cell surfaces. Heparan sulphate has marked sequence ersity that allows it to specifically interact with many proteins. This Review focuses on the multiple roles of heparan sulphate in inflammatory responses and, in particular, on its participation in almost every stage of leukocyte transmigration through the blood-vessel wall. Heparan sulphate is involved in the initial adhesion of leukocytes to the inflamed endothelium, the subsequent chemokine-mediated transmigration through the vessel wall and the establishment of both acute and chronic inflammatory reactions.

Immune deviation: A historical perspective

Publisher: Wiley

Date: 10-1996

DOI: 10.1038/ICB.1996.75

Abstract: In this paper I have reviewed my early studies, between 1966 and 1976, on the phenomenon of immune deviation. Initially summarized are experiments with different forms of the flagellin antigen from Salmonella adelaide which established the inverse relationship between delayed-type hypersensitivity (DTH) and antibody formation. Based on the flagellin studies, many of the key factors which determine whether an antigen will induce either DTH or antibody formation were delineated. These factors are just as relevant today as they were 25 years ago. Subsequent analyses at the cellular level demonstrated that different T cell subsets mediate DTH and T cell help and maintain immune deviation by suppressor mechanisms. A number of fundamental questions raised by this early work remain unanswered and are discussed. These include the nature of the primary signalling events which initiate immune deviation, the role of B cells in the deviating process and the mechanism by which CD8+T cells suppress antibody production.

Cell surface expression of the 300 kDa mannose-6-phosphate receptor by activated T lymphocytes

Publisher: Wiley

Date: 10-2001

DOI: 10.1046/J.1440-1711.2001.01026.X

Abstract: Phosphosugars, such as mannose-6-phosphate (M6P), have been shown previously to display anti-inflammatory properties, notably inhibition of experimental autoimmune encephalomyelitis (EAE) and adjuvant-induced arthritis in rats. It has been proposed that M6P exerts its anti-inflammatory effect by displacing lysosomal enzymes, which are involved in T-cell extravasation into inflammatory sites, from the 300 kDa mannose-6- phosphate receptor (MPR-300) on the surface of T cells. If this model is correct MPR-300 should be selectively expressed on the surface of activated T cells, as T cell entry into the central nervous system in EAE depends on the T cells being in an activated state. Thus, the present study examines whether cell surface expression of MPR-300 by T lymphocytes correlates with their state of activation and whether T cells in inflammatory sites express the receptor. Flow cytometric studies showed MPR-300 to be absent from the surface of unstimulated rat T cells isolated from peripheral blood and lymphoid tissues, and T cells resident within the peritoneal cavity. In contrast, MPR-300 was expressed on activated T cells derived from an inflammatory peritoneal exudate. In vitro studies demonstrated transient expression of MPR-300 on the surface of splenic T cells following stimulation with Con A. MPR-300 was also induced on T-cell lines by antigen stimulation. These data demonstrate that T cells in inflammatory sites express MPR-300 on their surface and activation of T lymphocytes induces cell surface expression of MPR-300. Such findings are consistent with the hypothesis that cell surface MPR-300 is required for the entry of T cells into inflammatory sites.

Evidence That Carrier-Specific B Cells Play a Role in Cellular Collaboration

Publisher: Wiley

Date: 03-1976

DOI: 10.1111/J.1365-3083.1976.TB00269.X

Abstract: Evidence is presented showing that carrier-specific B cells play a significant role in inducing an anti-hapten antibody response to the thymus-dependent antigen, dinitrophenylated monomeric flagellin (DNP-MON), both in vivo and in vitro. Inactivation of carrier-specific B cells by (125I) polymeric flagellin (POL) suicide decreased the anti-DNP IgM response by 40%-50% and reduced the anti-DNP IgG response by 85%-90%. Several lines of evidence established that the suicide procedure eliminated carrier-specific B cells rather than carrier-specific T (helper) cells.

Erratum: A functional heparan sulfate mimetic implicates both heparanase and heparan sulfate in tumor angiogenesis and invasion in a mouse model of multistage cancer (Oncogene (2005) 24 (4037-4051) DOI: 10.1038/sj.onc.1208602)

Publisher: Springer Science and Business Media LLC

Date: 09-06-2005

DOI: 10.1038/SJ.ONC.1208770

Immunology and Cell Biology publication of the year award 2008

Publisher: Wiley

Date: 10-2009

DOI: 10.1038/ICB.2009.58

Characterisation of the anticoagulant properties of a range of structurally diverse sulfated oligosaccharides

Publisher: Elsevier BV

Date: 08-2001

DOI: 10.1016/S0049-3848(01)00314-0

Abstract: In this study, 17 sulfated oligosaccharides were assessed by the activated partial thromboplastin time (APTT) test for their anticoagulant activity and nine were found to exhibit significant activity. Chain length, monosaccharide makeup, and linkage all appear to be critical factors in determining anticoagulant activity, with the most active compounds being five- to sixfold less potent than unfractionated heparin (UFH). Phosphomannopentaose sulfate (PI-88), one of the most active sulfated oligosaccharides and a promising anticancer drug, was selected for further study. PI-88 gave a more linear APTT dose-response curve and displayed less patient-to-patient variation than UFH, with its activity being neutralised by protamine sulfate. However, PI-88 showed considerable species-to-species variation in its anticoagulant effect. It was found that PI-88 acted as an anticoagulant by enhancing the ability of heparin cofactor II (HCII) to inhibit thrombin, and did not act via antithrombin III (AT-III) in either inhibiting Factor Xa or thrombin. PI-88 also mildly prolonged the prothrombin time (PT), whilst it had no platelet pro-aggregatory activity, nor did it demonstrate direct fibrinolytic activity. Thus, PI-88 represents a potential antithrombotic agent deserving further study.

Reduced retinal microvascular density, improved forepaw reach, comparative microarray and gene set enrichment analysis with c-jun targeting DNA enzyme

Publisher: Public Library of Science (PLoS)

Date: 17-07-2012

DOI: 10.1371/JOURNAL.PONE.0039160

Comparison of antigens recognized by xenogeneic and allogeneic anti-Ia antibodies: Evidence for two classes of Ia antigens

Publisher: Springer Science and Business Media LLC

Date: 12-1978

DOI: 10.1007/BF01563926

Cleavage of bacterial flagellin with proteolytic enzymes. I. Physicochemical and antigenic properties of the tryptic and peptic peptides

Publisher: Elsevier BV

Date: 02-1972

DOI: 10.1016/0019-2791(72)90036-5

Abstract: Uranium is toxic and radioactive traces of it can be found in natural water and soils. High concentrations of it in biological systems cause genetic disorders and diseases. For the in vivo diagnosis, micro and nano range detection limits are required. Here, an electrochemical assay for trace toxic uranium was searched using stripping voltammetry. Renewable and simplified graphite pencils electrode (PE) was used in a three-electrode cell system. Seawater was used instead of an electrolyte solution. This setup can yield good results and the detection limit was attained to be at 10 μgL(-1). The developed skill can be applied to organic liver cell.

The antiangiogenic properties of sulfated β-cyclodextrins in anticancer formulations incorporating 5-fluorouracil

Publisher: Ovid Technologies (Wolters Kluwer Health)

Date: 08-2013

DOI: 10.1097/CAD.0B013E3283626FDD

Monoclonal antibody detection of carbohydrate-defined and protein-defined H-2kK antigens

Publisher: Elsevier BV

Date: 07-1981

DOI: 10.1016/0161-5890(81)90038-9

Abstract: Whether types of hospitals with high readmission rates also have high overall postdischarge acute care utilization (including emergency department and observation care) is unknown. Cross-sectional analysis. Nonfederal United States acute care hospitals. Using methodology established by the Centers for Medicare & Medicaid Services, we calculated each hospital's "excess days in acute care" for fee-for-service (FFS) Medicare beneficiaries aged over 65 years discharged after hospitalization for acute myocardial infarction, heart failure (HF), or pneumonia, representing the mean difference between predicted and expected total days of acute care utilization in the 30 days following hospital discharge, per 100 discharges. We assessed the multivariable association of 8 hospital characteristics with excess days in acute care and the proportion of hospitals with each characteristic that were statistical outliers (95% credible interval estimate does not include 0). We included 2184 hospitals for acute myocardial infarction [228 (10.4%) better than expected, 549 (25.1%) worse than expected], 3720 hospitals for HF [484 (13.0%) better and 840 (22.6%) worse], and 4195 hospitals for pneumonia [673 (16.0%) better, 1005 (24.0%) worse]. Results for all conditions were similar. Worse than expected outliers for pneumonia included: 18.8% of safety net hospitals versus 26.1% of nonsafety net hospitals 16.7% of public hospitals versus 33.1% of for-profit hospitals 19.5% of nonteaching hospitals versus 52.2% of major teaching hospitals 7.9% of rural hospitals versus 42.1% of large urban hospitals 5.9% of hospitals with 24- 500 beds and 29.0% of hospitals with nurse-to-bed ratios >1.0-1.5 versus 21.7% of hospitals with ratios >2.0. Including emergency department and observation stays in measures of postdischarge utilization produces similar results as measuring only readmissions in that major teaching, urban and for-profit hospitals still perform disproportionately poorly versus nonteaching or public hospitals. However, it enables identification of more outliers and a more granular assessment of the association of hospital factors and outcomes.

Extracellular Matrix Components in the Pathogenesis of Type 1 Diabetes

Publisher: Springer Science and Business Media LLC

Date: 26-10-2014

DOI: 10.1007/S11892-014-0552-7

The fate of self-reactive B cells depends primarily on the degree of antigen receptor engagement and availability of T cell help

Publisher: Rockefeller University Press

Date: 05-1996

DOI: 10.1084/JEM.183.5.2313

Abstract: Self-reactive B cells from tolerant double-transgenic (Dbl-Tg) mice coexpressing hen egg lysozyme (HEL) and rearranged anti-HEL immunoglobulin genes have a relatively short life span when compared to normal B cells, irrespective of whether they are exposed to antigen in multivalent membrane-bound form (mHEL-Dbl-Tg mice) or soluble form (sHEL-Dbl-Tg mice). The factors responsible for determining the fate of these B cells after encounter with self-antigen were investigated using a cell-tracking technique in which anti-HEL Ig-Tg spleen cells were labeled with the intracellular dye 5-carboxyfluorescein diacetate-succinimidyl ester (CFSE) and injected either into non-Tg recipients or a variety of HEL-Tg hosts. In non-Tg recipients, HEL-binding B cells persisted in the circulation and could be detected in the follicles of the spleen for at least 5 d. On transfer into either mHEL-Tg or sHEL-Tg hosts, they underwent activation and then rapidly disappeared from the blood and spleen over the next 3 d, consistent with the short life span reported previously. Immunohistology of spleens from sHEL-Tg recipients indicated that the transferred B cells had migrated to the outer margins of the periarteriolar lymphoid sheath (PALS), where they were detectable for 24 h before being lost. The positioning of B cells in the outer PALS depended on a critical threshold of Ig receptor binding corresponding to a serum HEL concentration between 0.5 and 15 ng/ml, but was not restricted to endogenously expressed HEL in that the same migratory pattern was observed after transfer into non-Tg recipients given exogenous (foreign) HEL. Moreover, bone marrow-derived immature Ig-Tg B cells homed to the outer PALS of sHEL-Tg mice and then disappeared at the same rate as mature B cells, indicating that the stage of maturation did not influence the fate of self-reactive B cells in a tolerant environment. On the other hand, HEL-binding B cells transferred into sHEL-Dbl-Tg recipients persisted over the 3-d period of study, apparently due to insufficient availability of antigen, as indicated by the fact that the degree of Ig receptor downregulation on the transferred B cells was much less than in sHEL-Tg recipients. If T cell help was provided to Ig-Tg B cells at the time of transfer into sHEL-Tg recipients in the form of preactivated CD4+ T cells specific for major histocompatibility complex-peptide complexes on the B cell surface, HEL-binding B cells migrated through the outer PALS of the spleen to the follicle, where they formed germinal centers, or to adjacent red pulp, where they formed proliferative foci and secreted significant amounts of anti-HEL antibody. Taken together, these results indicated that the outcome of the interaction between self-antigen and B cells is largely determined by a combination of the degree of receptor engagement and availability of T cell help.

New fluorescent dyes for lymphocyte migration studies

Publisher: Elsevier BV

Date: 10-1990

DOI: 10.1016/0022-1759(90)90322-M

Abstract: 16 fluorochromes were examined for their ability to label viable lymphocytes in vitro and yield fluorescence detectable by fluorescence microscopy and flow cytometry. Of these fluorochromes, four intracellular dyes were found to be suitable for in vivo migration studies. They were H33342, the well known DNA-binding dye which excites and emits in the UV range, and three fluorescein based cytoplasmic dyes, namely BCECF-AM, Calcein-AM and CFSE which excite and emit in the visible range. Lymphocytes labelled with H33342, BCECF-AM and Calcein-AM were suitable for short term in vivo migration experiments with detection by flow cytometry 2-3 days post injection. In contrast lymphocytes labelled with CFSE, a fluorochrome which can covalently couple with intracellular macromolecules, were detected by flow cytometry up to 8 weeks post injection and thus this fluorochrome is ideal for long term migration experiments. Due to marked differences in fluorescence profiles, BCECF-AM and Calcein-AM could be used for short term double labelling experiments using the flow cytometer in which entry of injected lymphocytes into lymphoid organs was quantified. Similarly, in vivo localization of lymphocyte subpopulations could be examined by fluorescence microscopy utilizing differences in fluorescence excitation and emission spectra of lymphocytes labelled with H33342 and one of the fluorescein based dyes.

A Comparison of the Terminal Protein and Hexon Polypeptides of Avian and Human Adenoviruses

Publisher: Microbiology Society

Date: 06-1983

DOI: 10.1099/0022-1317-64-6-1375

Abstract: It was found that the virion terminal protein of chick embryo lethal orphan (CELO) virus had a molecular weight of 46,000, and the hexon a molecular weight of 100,000. 125I-labelled tryptic and chymotryptic peptide maps of the hexons and terminal proteins from CELO virus and human adenovirus type 5 (Ad5) differed. However, limited proteolysis of CELO virus and Ad5 terminal proteins by protease V8 showed similarities which were not detected in the case of the two hexons.

Inhibition of secondary IgG responses by N-acetyl-D-galactosamine.

Publisher: Wiley

Date: 1981

DOI: 10.1002/EJI.1830110305

Abstract: A range of monosaccharides were tested for their ability to inhibit a variety of in vitro immune response. The most striking specific inhibition was produced by N-acetyl-D-galactosamine (GalNAc). This sugar strongly inhibited the secondary IgG antibody response to two different hapten-carrier systems, but had no effect on primary and secondary IgM responses, generation of cytotoxic T cells to alloantigens and mixed lymphocyte reactions. By exposing secondary antibody cultures to GalNAc for varying periods of time, it was observed that GalNAc only exerted its inhibitory effect on day 4 of the culture, the day when IgG plaque-forming cells first appeared. Furthermore, GalNAc could override the action of T helper factor in T cell-depleted cultures. Collectively, these data indicate that GalNAc inhibits the initiation of IgG synthesis probably by blocking the interaction of a helper factor for IgG synthesis with its target cell.

The lymphocyte surface. III. Function of Fc receptor, C'3 receptor and surface Ig bearing lymphocytes: identification of a radioresistant B cell

Publisher: The Royal Society

Date: 19-11-1974

DOI: 10.1098/RSPB.1974.0083

Abstract: Rat thoracic duct lymphocytes (TDL) were depleted of Fc receptor (FcRL), C'3 receptor (CRL) or surface Ig bearing (IgL) lymphocytes as described in the preceding paper (Parish & Hayward 1974 b , part II). The FcRL, CRL and IgL populations were also recovered in pure form and their ability to transfer antibody responsiveness to irradiated recipients was assessed. It was found that TDL from rats primed to DNP, when depleted of IgL, lost their capacity to transfer a secondary 7S antibody response to DNP in contrast, depletion of FcRL did not affect the 7S response. CRL depletion eliminated the transferred primary 7S antibody response to polymerized flagellin and also reduced but did not eliminate the secondary 7S antibody response to DNP. Cells which are Ig + CR + appear to be the main source of secondary 7S precursors in the rat but Ig + CR - cells can also, under certain circumstances, serve the same function. CRL, IgL and FcRL purified from DNP-primed TDL were unable to transfer secondary responsiveness to DNP. However, the addition of CRL or IgL to TDL depleted of these subpopulations resulted in a reconstitution of the secondary 7S antibody response. By using cytotoxic alloantisera CRL were identified as 7S antibody precursors. In order to mount a 7S antibody response to DNP, CRLs required the collaboration of non-CRLs, the CRL population containing the 7S precursors and the non-CRL population contributing helper (T) cells. Removal of either CRL, IgL or FcRL from TDL had no effect on the 19S anti-DNP response which arose in the irradiated recipients. The same result was obtained whether the 19S and 7S antibodies were characterized by their 2ME sensitivity, by their sedimentation behaviour on sucrose gradients or by their expression of direct and indirect PFC. However, no conclusions could be drawn from these observations about the nature of 19S precursor cells because it was found that virtually all the cells producing 19S anti-DNP antibody originated from the irradiated hosts, indicating that radioresistant 19S precursors exist in the rat. On some occasions radioresistant 7S precursors were also detected. To be activated by antigen both radioresistant populations require the presence of primed helper (T) cells. These results emphasize the extreme caution required in interpreting experiments that involve cell transfers into irradiated recipients.

A possible aberrant line of BALB/c-H-2dm2 mutant mice.

Publisher: The American Association of Immunologists

Date: 11-1982

DOI: 10.4049/JIMMUNOL.129.5.2314.A

University Of Melbourne

Location: Australia

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Australian National University

Location: Australia

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Vascular Biology

Start Date: 2002

End Date: 2006

Funder: National Health and Medical Research Council

Regional Facility For Real Time Analysis Of Molecular Interactions

Start Date: 2009

End Date: 2009

Funder: Australian Research Council

Vascular Biology

Start Date: 2007

End Date: 2011

Funder: National Health and Medical Research Council

Heparin-induced Thrombocytopenia And Thrombosis: Better Understanding Of Pathogenesis And Improving Diagnosis And Treatment

Start Date: 2012

End Date: 2014

Funder: National Health and Medical Research Council

Multiphoton Confocal Microscope

Start Date: 2010

End Date: 2010

Funder: Australian Research Council

Novel Mechanisms, Molecular Targets And Therapies In Cardiovascular Disease

Start Date: 2014

End Date: 2020

Funder: National Health and Medical Research Council

Centre For Integrative Legume Research

Start Date: 2003

End Date: 2010

Funder: Australian Research Council

State-of-the-art Facility For Human And Animal Virus Research In The Canberra And Surrounding Regions

Start Date: 2009

End Date: 2009

Funder: Australian Research Council

Linkage Infrastructure, Equipment And Facilities - Grant ID: LE130100210

Start Date: 2013

End Date: 12-2013

Amount: $350,000.00

Funder: Australian Research Council

Linkage Projects - Grant ID: LP110100680

Start Date: 03-2011

End Date: 08-2015

Amount: $105,000.00

Funder: Australian Research Council

Discovery Projects - Grant ID: DP1096299

Start Date: 02-2010

End Date: 02-2013

Amount: $390,000.00

Funder: Australian Research Council

Linkage Infrastructure, Equipment And Facilities - Grant ID: LE0454209

Start Date: 03-2004

End Date: 06-2005

Amount: $126,326.00

Funder: Australian Research Council

ARC Centres Of Excellence - Grant ID: CE0348212

Start Date: 2003

End Date: 12-2011

Amount: $16,900,000.00

Funder: Australian Research Council

Linkage Infrastructure, Equipment And Facilities - Grant ID: LE100100078

Start Date: 2010

End Date: 12-2010

Amount: $600,000.00

Funder: Australian Research Council

Linkage Infrastructure, Equipment And Facilities - Grant ID: LE0989077

Start Date: 2009

End Date: 12-2009

Amount: $225,600.00

Funder: Australian Research Council

Linkage Infrastructure, Equipment And Facilities - Grant ID: LE0989564

Start Date: 06-2009

End Date: 06-2010

Amount: $150,000.00

Funder: Australian Research Council