John Hayball

Maternal host responses to poly(I:C) during pregnancy leads to both dysfunctional immune profiles and altered behaviour in the offspring

Publisher: Wiley

Date: 26-05-2020

DOI: 10.1111/AJI.13260

Dasatinib inhibits the secretion of TNF-a following TLR stimulation in vitro and in vivo

Publisher: Elsevier BV

Date: 12-2009

DOI: 10.1016/J.EXPHEM.2009.09.007

Abstract: Dasatinib (SPRYCEL, BMS-354825) is a small molecule Src/Abl tyrosine kinase inhibitor approved for the treatment of chronic myeloid leukemia and Philadelphia chromosome-positive acute lymphoblastic leukemia. However, kinases inhibited by dasatinib are also involved in the induction and regulation of innate immunity. The purpose of this study was to evaluate the effect of dasatinib on cytokine secretion in response to toll-like receptor (TLR) stimulation. Dasatinib-treated mice were administered intraperitoneally with lipopolysaccharide (LPS) and serum cytokine (tumor necrosis factor-alpha [TNF-alpha], interleukin [IL]-10, and IL-6) levels and neutrophil accumulation in the lungs were analyzed. Cytokine secretions (TNF-alpha and IL-6) from TLR3-, TLR4-, and TLR9-stimulated RAW264.7, as well as TLR4- and TLR9-stimulated bone marrow-derived macrophages (BMDM) were also evaluated. Dasatinib-treated mice had reduced serum levels of TNF-alpha in response to LPS administration however, other inflammatory hallmarks of systemic LPS administration, such as secretion of IL-6 and accumulation of neutrophils in the lung, were unaffected. In contrast to the reduced TNF-alpha levels, dasatinib treatment increased serum levels of IL-10 following LPS administration. The production of TNF-alpha was also impaired in vitro in response to TLR3, TLR4, and TLR9 stimulation of the mouse macrophage cell line RAW264.7, as well as TLR4 and TLR9 stimulation of BMDM IL-6 production was also impaired in dasatinib-treated BMDM. These findings further support the ability of dasatinib to modulate the host immune response and highlights scope for off-target applications of dasatinib for the control of TNF-alpha-mediated inflammatory disorders.

MHC multimerization, antigen expression and the induction of APC amnesia in the developing immune response

Publisher: Wiley

Date: 02-1999

DOI: 10.1046/J.1440-1711.1999.00794.X

Abstract: Class II multimer formation is an important event in immune recognition. Not only is multimerization a prerequisite for T cell activation, but it is a signal to APC. In the present article, we propose that multimerization can result in the specific removal of ligand complexes from the cell surface of the APC, an event which may influence the overall pattern of T cell reactivity.

Gamma-Irradiated Influenza Virus Uniquely Induces IFN-I Mediated Lymphocyte Activation Independent of the TLR7/MyD88 Pathway

Publisher: Public Library of Science (PLoS)

Date: 05-10-2011

DOI: 10.1371/JOURNAL.PONE.0025765

The vaccinia virus based Sementis Copenhagen Vector vaccine against Zika and chikungunya is immunogenic in non-human primates

Publisher: Springer Science and Business Media LLC

Date: 02-06-2020

DOI: 10.1038/S41541-020-0191-8

Abstract: The Sementis Copenhagen Vector (SCV) is a new vaccinia virus-derived, multiplication-defective, vaccine technology assessed herein in non-human primates. Indian rhesus macaques ( Macaca mulatta ) were vaccinated with a multi-pathogen recombinant SCV vaccine encoding the structural polyproteins of both Zika virus (ZIKV) and chikungunya virus (CHIKV). After one vaccination, neutralising antibody responses to ZIKV and four strains of CHIKV, representative of distinct viral genotypes, were generated. A second vaccination resulted in significant boosting of neutralising antibody responses to ZIKV and CHIKV. Following challenge with ZIKV, SCV-ZIKA/CHIK-vaccinated animals showed significant reductions in viremias compared with animals that had received a control SCV vaccine. Two SCV vaccinations also generated neutralising and IgG ELISA antibody responses to vaccinia virus. These results demonstrate effective induction of immunity in non-human primates by a recombinant SCV vaccine and illustrates the utility of SCV as a multi-disease vaccine platform capable of delivering multiple large immunogens.

Biomaterial Surface Hydrophobicity-Mediated Serum Protein Adsorption and Immune Responses

Publisher: American Chemical Society (ACS)

Date: 16-07-2019

DOI: 10.1021/ACSAMI.9B09900

Abstract: The nature of the protein corona forming on biomaterial surfaces can affect the performance of implanted devices. This study investigated the role of surface chemistry and wettability on human serum-derived protein corona formation on biomaterial surfaces and the subsequent effects on the cellular innate immune response. Plasma polymerization, a substrate-independent technique, was employed to create nanothin coatings with four specific chemical functionalities and a spectrum of surface charges and wettability. The amount and type of protein adsorbed was strongly influenced by surface chemistry and wettability but did not show any dependence on surface charge. An enhanced adsorption of the dysopsonin albumin was observed on hydrophilic carboxyl surfaces while high opsonin IgG2 adsorption was seen on hydrophobic hydrocarbon surfaces. This in turn led to a distinct immune response from macrophages hydrophilic surfaces drove greater expression of anti-inflammatory cytokines by macrophages, whilst surface hydrophobicity caused increased production of proinflammatory signaling molecules. These findings map out a unique relationship between surface chemistry, hydrophobicity, protein corona formation, and subsequent cellular innate immune responses the potential outcomes of these studies may be employed to tailor biomaterial surface modifications, to modulate serum protein adsorption and to achieve the desirable innate immune response to implanted biomaterials and devices.

Creating Nano-engineered Biomaterials with Well-Defined Surface Descriptors

Publisher: American Chemical Society (ACS)

Date: 16-05-2018

DOI: 10.1021/ACSANM.8B00458

In vivo targeting of dead tumor cells in a murine tumor model using a monoclonal antibody specific for the La autoantigen

Publisher: American Association for Cancer Research (AACR)

Date: 15-09-2007

DOI: 10.1158/1078-0432.CCR-07-0964

Abstract: Purpose: To investigate the potential of the La-specific monoclonal antibody (mAb) 3B9 as an in vivo tumor-targeting agent. Experimental Design: The murine EL4 lymphoma cell line was used for in vitro studies and the EL4 model in which apoptosis was induced with cyclophosphamide and etoposide was used for in vivo studies. In vitro studies compared 3B9 binding in the EL4 cell with that in its counterpart primary cell type of the thymocyte. For in vivo studies, 3B9 was intrinsically or extrinsically labeled with carbon-14 or 1,4,7,10-tetra-azacylododecane-N,N′,N″,N″″-tetraacetic acid–indium-111, respectively, and biodistribution of the radiotracers was investigated in EL4 tumor-bearing mice, which were treated or not with chemotherapy. Results: La-specific 3B9 mAb bound EL4 cells rather than thymocytes, and binding was detergent resistant. 3B9 binding to dead EL4 cells in vitro was specific, rapid, and saturable. Significantly, more 3B9 bound dead EL4 tumor explant cells after host mice were treated with chemotherapy, which suggested that DNA damage induced 3B9 binding. Tumor binding of 3B9 in vivo was antigen specific and increased significantly after chemotherapy. Tumor accumulation of 3B9 peaked at ∼50% of the injected dose per gram of tumor 72 h after chemotherapy and correlated with increased tumor cell death. Tumor/organ ratios of 3B9 biodistribution, which included the tumor/blood ratio, exceeded unity 48 or more hours after chemotherapy. Conclusions: La-specific mAb selectively targeted dead tumor cells in vivo, and targeting was augmented by cytotoxic chemotherapy. This novel cell death radioligand may be useful both for radioimmunoscintigraphy and radioimmunotherapy.

Recombinant fowlpox virus elicits transient cytotoxic T cell responses due to suboptimal innate recognition and recruitment of T cell help

Publisher: Elsevier BV

Date: 07-2008

DOI: 10.1016/J.VACCINE.2008.05.015

Abstract: Recombinant fowlpox viruses (FPVs) have been used in a variety of vaccine strategies however strong data clearly demonstrating the characteristics of the strength and nature of the resultant immune response elicited by these vectors are lacking. By utilising a recombinant variant of FPV which expresses the nominal antigen chicken ovalbumin (OVA), and assessing innate FPV- and OVA-specific adaptive immune responses, we show that recombinant FPV induces a rapid type I interferon (IFN) response, mediated primarily by plasmacytoid dendritic cells (pDCs). These cells are necessary for the development of a strong but transient CD8(+) T cell effector response directed against OVA-expressing target cells. We propose that a combination of suboptimal type I IFN production, poor CD4(+) T cell helper function and inefficient DC licensing likely contribute to this transient response. These findings now provide a sound basis for rational modifications to be made to recombinant FPV, designed to improve subsequent vaccine responses.

A Practical Guide to Prepare and Synthetically Modify Graphene Quantum Dots

Publisher: Wiley

Date: 07-02-2019

DOI: 10.1002/ADFM.201808740

Peripheral Interleukin-1β Levels are Elevated in Chronic Tension-Type Headache Patients

Publisher: Hindawi Limited

Date: 2013

DOI: 10.1155/2013/796161

Abstract: BACKGROUND: Tension-type headache is the most common form of headache and its chronic form, chronic tension-type headache (CTTH), is one of the most difficult to treat. The etiology of CTTH is not well understood, but is believed to be multifactorial and to vary among in iduals. In the present study, the authors sought to identify common mechanisms of CTTH pathology. Empirical studies have implicated various immunomodulatory cytokines as mediators of chronic pain disorders, including CTTH. OBJECTIVES: To determine the role of peripheral cytokines and genetic factors in the development of CTTH. METHODS: A panel of cytokines hypothesized to play a role in the pathogenesis of CTTH was measured using cytometric bead arrays and ELISAs in 56 in iduals with CTTH and 42 healthy control participants between 18 and 65 years of age. RESULTS: Levels of interleukin (IL)-1β were significantly elevated in participants diagnosed with CTTH relative to healthy controls, while IL-18 levels were found to be significantly elevated in men with CTTH. Because the levels of these immune mediators were increased in the apparent absence of injury or infection, the authors sought to determine whether genetic changes were responsible for fluctuations in cytokine levels. Polymerase chain reaction and restriction fragment length polymorphism analyses were used to determine in idual genotypes at key single nucleotide polymorphism positions in the IL-1B gene. No association was observed between CTTH and single nucleotide polymorphisms in the IL-1β gene. CONCLUSIONS: These findings suggest that increases in key proinflammatory cytokine levels are associated with CTTH and the pathology of the disorder involves sterile neurovascular inflammation.

An Empirical Approach towards the Efficient and Optimal Production of Influenza-Neutralizing Ovine Polyclonal Antibodies Demonstrates That the Novel Adjuvant CoVaccine HT™ Is Functionally Superior to

Publisher: Public Library of Science (PLoS)

Date: 23-07-2013

DOI: 10.1371/JOURNAL.PONE.0068895

Production of a Chikungunya Vaccine Using a CHO Cell and Attenuated Viral-Based Platform Technology

Publisher: Elsevier BV

Date: 10-2017

DOI: 10.1016/J.YMTHE.2017.06.017

Doxorubicin-Loaded Delta Inulin Conjugates for Controlled and Targeted Drug Delivery: Development, Characterization, and In Vitro Evaluation

Publisher: MDPI AG

Date: 06-11-2019

DOI: 10.3390/PHARMACEUTICS11110581

Abstract: Delta inulin, also known as microparticulate inulin (MPI), was modified by covalently attaching doxorubicin to its nanostructured surface for use as a targeted drug delivery vehicle. MPI is readily endocytosed by monocytes, macrophages, and dendritic cells and in this study, we sought to utilize this property to develop a system to target anti-cancer drugs to lymphoid organs. We investigated, therefore, whether MPI could be used as a vehicle to deliver doxorubicin selectively, thereby reducing the toxicity of this antibiotic anthracycline drug. Doxorubicin was covalently attached to the surface of MPI using an acid–labile linkage to enable pH-controlled release. The MPI-doxorubicin conjugate was characterized using FTIR and SEM, confirming covalent attachment and indicating doxorubicin coupling had no obvious impact on the physical nanostructure, integrity, and cellular uptake of the MPI particles. To simulate the stability of the MPI-doxorubicin in vivo, it was stored in artificial lysosomal fluid (ALF, pH 4.5). Although the MPI-doxorubicin particles were still visible after 165 days in ALF, 53% of glycosidic bonds in the inulin particles were hydrolyzed within 12 days in ALF, reflected by the release of free glucose into solution. By contrast, the fructosidic bonds were much more stable. Drug release studies of the MPI-doxorubicin in vitro, demonstrated a successful pH-dependent controlled release effect. Confocal laser scanning microscopy studies and flow cytometric analysis confirmed that when incubated with live cells, MPI-doxorubicin was efficiently internalized by immune cells. An assay of cell metabolic activity demonstrated that the MPI carrier alone had no toxic effects on RAW 264.7 murine monocyte/macrophage-like cells, but exhibited anti-cancer effects against HCT116 human colon cancer cells. MPI-doxorubicin had a greater anti-cancer cell effect than free doxorubicin, particularly when at lower concentrations, suggesting a drug-sparing effect. This study establishes that MPI can be successfully modified with doxorubicin for chemotherapeutic drug delivery.

The utility of monitoring peripheral blood lymphocyte subsets by flow cytometric analysis in patients with rheumatological diseases treated with rituximab

Publisher: Elsevier BV

Date: 05-2017

DOI: 10.1016/J.AUTREV.2017.03.003

IDENTIFICATION OF 2 BINDING-SITES IN STAPHYLOCOCCAL ENTEROTOXIN-B THAT CONFER SPECIFICITY FOR TCR V(BETA) GENE-PRODUCTS

Publisher: Oxford University Press (OUP)

Date: 1994

DOI: 10.1093/INTIMM/6.2.199

Abstract: The enterotoxins produced by Staphylococcus aureus are potent mitogens. They stimulate T cells in an oligoclonal fashion that is dependent on the expression of particular variable region gene elements in the beta-chain of the TCR (V beta). The fourth hypervariable loop of the TCR beta-chain is generally regarded as the site of contact for both viral and microbial superantigens. Recently, residues 60 and 61 of staphylococcal enterotoxin B (SEB) have been highlighted as central to the interaction of this toxin with the TCR. We have, therefore, analysed a series of toxins with mutations at these positions to investigate how amino acid substitutions affect the ability of mutant toxins to stimulate both human and mouse T cells. Each of the variant toxins induced proliferation in a murine V beta 8.3 T cell clone, whereas a V beta 8.1 T cell clone only responded to native toxin. A panel of nine human T cell clones expressing six different V beta elements, all of which responded to native SEB, was tested for reactivity to the variant toxins. Only one V beta 19.1+ T cell clone was found to be sensitive to substitution at positions 60 and 61 in a manner analogous to the murine V beta 8.1 T cell clone. Semi-quantitative analysis of the TCR V beta expression of human T cell lines expanded with native and mutant SEB revealed that none of the variant toxins could stimulate T cells that expressed V beta 19.1. Taken together, these results suggest that the interaction of mouse V beta 8.1 and human V beta 19.1 TCRs with SEB differs from other TCRs. Sequence comparisons of the different TCR V beta chains indicated that residues in the second complementarity determining region (CDR2) interact with the 60-61 loop of SEB. Therefore, a minimum of two distinct binding modules confer specificity to the interaction of the TCR with SEB.

Serrulatane Diterpenoid from Eremophila neglecta Exhibits Bacterial Biofilm Dispersion and Inhibits Release of Pro-inflammatory Cytokines from Activated Macrophages

Publisher: American Chemical Society (ACS)

Date: 04-12-2015

DOI: 10.1021/ACS.JNATPROD.5B00833

Abstract: The purpose of this study was to assess the biofilm-removing efficacy and inflammatory activity of a serrulatane diterpenoid, 8-hydroxyserrulat-14-en-19-oic acid (1), isolated from the Australian medicinal plant Eremophila neglecta. Biofilm breakup activity of compound 1 on established Staphylococcus epidermidis and Staphylococcus aureus biofilms was compared to the antiseptic chlorhexidine and antibiotic levofloxacin. In a time-course study, 1 was deposited onto polypropylene mesh to mimic a wound dressing and tested for biofilm removal. The ex-vivo cytotoxicity and effect on lipopolysaccharide-induced pro-inflammatory cytokine release were studied in mouse primary bone-marrow-derived macrophage (BMDM) cells. Compound 1 was effective in dispersing 12 h pre-established biofilms with a 7 log10 reduction of viable bacterial cell counts, but was less active against 24 h biofilms (approximately 2 log10 reduction). Compound-loaded mesh showed dosage-dependent biofilm-removing capability. In addition, compound 1 displayed a significant inhibitory effect on tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) secretion from BMDM cells, but interleukin-1 beta (IL-1β) secretion was not significant. The compound was not cytotoxic to BMDM cells at concentrations effective in removing biofilm and lowering cytokine release. These findings highlight the potential of this serrulatane diterpenoid to be further developed for applications in wound management.

Cytolytic DNA vaccine encoding lytic perforin augments the maturation of - and antigen presentation by - dendritic cells in a time-dependent manner

Publisher: Springer Science and Business Media LLC

Date: 17-08-2017

DOI: 10.1038/S41598-017-08063-1

Abstract: The use of cost-effective vaccines capable of inducing robust CD8 + T cell immunity will contribute significantly towards the elimination of persistent viral infections and cancers worldwide. We have previously reported that a cytolytic DNA vaccine encoding an immunogen and a truncated mouse perforin (PRF) protein significantly augments anti-viral T cell (including CD8 + T cell) immunity. Thus, the current study investigated whether this vaccine enhances activation of dendritic cells (DCs) resulting in greater priming of CD8 + T cell immunity. In vitro data showed that transfection of HEK293T cells with the cytolytic DNA resulted in the release of lactate dehydrogenase, indicative of necrotic/lytic cell death. In vitro exposure of this lytic cell debris to purified DCs from naïve C57BL/6 mice resulted in maturation of DCs as determined by up-regulation of CD80/CD86. Using activation roliferation of adoptively transferred OT-I CD8 + T cells to measure antigen presentation by DCs in vivo , it was determined that cytolytic DNA immunisation resulted in a time-dependent increase in the proliferation of OT-I CD8 + T cells compared to canonical DNA immunisation. Overall, the data suggest that the cytolytic DNA vaccine increases the activity of DCs which has important implications for the design of DNA vaccines to improve their translational prospects.

PD-L1 Is Not Constitutively Expressed on Tasmanian Devil Facial Tumor Cells but Is Strongly Upregulated in Response to IFN-γ and Can Be Expressed in the Tumor Microenvironment

Publisher: Frontiers Media SA

Date: 09-12-2016

DOI: 10.3389/FIMMU.2016.00581

Dasatinib inhibits recombinant viral antigen-specific murine CD4+ and CD8+ T-cell responses and NK-cell cytolytic activity in vitro and in vivo

Publisher: Elsevier BV

Date: 02-2009

DOI: 10.1016/J.EXPHEM.2008.09.013

Abstract: Dasatinib (BMS-354825) is a small molecule Src/Abl tyrosine kinase inhibitor approved for the treatment of chronic myeloid leukemia and Philadelphia chromosome-positive acute lymphoblastic leukemia. Members of the Src family of kinases are involved in the induction of innate and adaptive immunity. The purpose of this study was to evaluate the inhibitory action of dasatinib on antigen-specific CD8(+) and CD4(+) T-cell function, as well as natural killer (NK) cell cytotoxicity. To assess dasatinib-mediated inhibition of antigen-specific T-cell proliferation, transgenic CD4(+) and CD8(+) T cells specific for ovalbumin were utilized. Endogenous CD4(+) and CD8(+) T-cell responses were determined following immunization of dasatinib-treated or control mice with a nonreplicating recombinant virus. Clearance of the RMA-S cells, a major histocompatibility complex (MHC) class I-deficient thymoma sensitive to NK-cell lysis, was analyzed in mice undergoing dasatinib treatment. Dasatinib inhibited antigen-specific proliferation of murine CD4(+) and CD8(+) transgenic T cells in vitro and in vivo. Endogenous antigen-specific helper T-cell recall responses and induction of T-cell-mediated cytotoxicity following immunization with a nonreplicating recombinant virus were also inhibited. So to was the ability of NK cells to eliminate MHC class I-deficient cells in vivo. These findings suggest that dasatinib has the potential to modulate the host immune response at clinical doses and highlights scope for off target applications, e.g., therapeutic immunosuppression in the context of autoimmune pathogenesis and allogeneic tissue transplantation.

Toll-like receptor 7 gene deficiency and early-life Pneumovirus infection interact to predispose toward the development of asthma-like pathology in mice

Publisher: Elsevier BV

Date: 05-2013

DOI: 10.1016/J.JACI.2013.02.041

Abstract: Respiratory tract viruses are a major environmental risk factor for both the inception and exacerbations of asthma. Genetic defects in Toll-like receptor (TLR) 7-mediated signaling, impaired type I interferon responses, or both have been reported in asthmatic patients, although their contribution to the onset and exacerbation of asthma remains poorly understood. We sought to determine whether Pneumovirus infection in the absence of TLR7 predisposes to bronchiolitis and the inception of asthma. Wild-type and TLR7-deficient (TLR7(-/-)) mice were inoculated with the rodent-specific pathogen pneumonia virus of mice at 1 (primary), 7 (secondary), and 13 (tertiary) weeks of age, and pathologic features of bronchiolitis or asthma were assessed. In some experiments infected mice were exposed to low-dose cockroach antigen. TLR7 deficiency increased viral load in the airway epithelium, which became sloughed and necrotic, and promoted an IFN-α/β(low), IL-12p70(low), IL-1β(high), IL-25(high), and IL-33(high) cytokine microenvironment that was associated with the recruitment of type 2 innate lymphoid cells/nuocytes and increased TH2-type cytokine production. Viral challenge of TLR7(-/-) mice induced all of the cardinal pathophysiologic features of asthma, including tissue eosinophilia, mast cell hyperplasia, IgE production, airway smooth muscle alterations, and airways hyperreactivity in a memory CD4(+) T cell-dependent manner. Importantly, infections with pneumonia virus of mice promoted allergic sensitization to inhaled cockroach antigen in the absence but not the presence of TLR7. TLR7 gene defects and Pneumovirus infection interact to establish an aberrant adaptive response that might underlie virus-induced asthma exacerbations in later life.

Assessment of snapper (Pagrus auratus) natural IgM binding to bromelain treated sheep erythrocytes

Publisher: Elsevier BV

Date: 2005

DOI: 10.1016/J.FSI.2004.06.006

Extended boiling of peanut progressively reduces IgE allergenicity while retaining T cell reactivity

Publisher: Wiley

Date: 12-05-2016

DOI: 10.1111/CEA.12740

Abstract: Current peanut oral immunotherapy is h ered by frequent adverse events. It has been shown that boiling can reduce peanut allergenicity. Hypoallergenic peanut products have the potential to reduce treatment-related reactions during desensitization. To show that extended boiling (for up to 12 h) can progressively reduce peanut allergenicity while retaining T cell reactivity. Raw peanuts were boiled for half, 1, 2, 4 and 12 h in deionized water. After dehydration, boiled and raw peanuts were ground, defatted and soluble proteins extracted in PBS and cooking water (leachate) retained. SDS-PAGE, Western blot, inhibition ELISA, mass spectrometry and skin prick test were used to characterize changes to peanut allergens and human IgE reactivity. T cell responses to raw and boiled peanut extracts were determined by proliferation of CD4+/CD25+/CD134+ T cells in peanut-allergic and non-allergic in iduals. Extended boiling progressively reduced peanut allergenicity through a combination of leaching of allergens into cooking water, fragmentation of allergens and denaturation of conformational epitopes. Two-hour boiling led to an eightfold reduction in IgE binding capacity of boiled peanuts as determined by inhibition ELISA, while 12-h boiling led to a 19-fold reduction. Mass spectrometry revealed an increasing number of unique allergen peptides with longer boiling times. Raw, 2- and 12-h boiled peanut extracts were equivalent in their ability to stimulate T cell activation and proliferation. Progressive reduction in peanut allergenicity with extended boiling does not affect T cell reactivity. Boiled peanuts may be a candidate for oral immunotherapy.

Arthritogenic Alphavirus Vaccines: Serogrouping Versus Cross-Protection in Mouse Models

Publisher: MDPI AG

Date: 05-05-2020

DOI: 10.3390/VACCINES8020209

Abstract: Chikungunya virus (CHIKV), Ross River virus (RRV), o’nyong nyong virus (ONNV), Mayaro virus (MAYV) and Getah virus (GETV) represent arthritogenic alphaviruses belonging to the Semliki Forest virus antigenic complex. Antibodies raised against one of these viruses can cross-react with other serogroup members, suggesting that, for instance, a CHIKV vaccine (deemed commercially viable) might provide cross-protection against antigenically related alphaviruses. Herein we use human alphavirus isolates (including a new human RRV isolate) and wild-type mice to explore whether infection with one virus leads to cross-protection against viremia after challenge with other members of the antigenic complex. Persistently infected Rag1-/- mice were also used to assess the cross-protective capacity of convalescent CHIKV serum. We also assessed the ability of a recombinant poxvirus-based CHIKV vaccine and a commercially available formalin-fixed, whole-virus GETV vaccine to induce cross-protective responses. Although cross-protection and/or cross-reactivity were clearly evident, they were not universal and were often suboptimal. Even for the more closely related viruses (e.g., CHIKV and ONNV, or RRV and GETV), vaccine-mediated neutralization and/or protection against the intended homologous target was significantly more effective than cross-neutralization and/or cross-protection against the heterologous virus. Effective vaccine-mediated cross-protection would thus likely require a higher dose and/or more vaccinations, which is likely to be unattractive to regulators and vaccine manufacturers.

CD4+ T cells cross-compete for MHC class II-restricted peptide antigen complexes on the surface of antigen presenting cells

Publisher: Wiley

Date: 04-2004

DOI: 10.1046/J.0818-9641.2004.01233.X

Scrutinizing calcium flux oscillations in T lymphocytes to deduce the strength of stimulus

Publisher: Springer Science and Business Media LLC

Date: 14-01-2015

DOI: 10.1038/SREP07760

Evaluation of trained immunity by β‐1, 3 (d)‐glucan on murine monocytes in vitro and duration of response in vivo

Publisher: Wiley

Date: 14-03-2017

DOI: 10.1038/ICB.2017.13

Cortistatin binds to TNF-α receptors and protects against osteoarthritis

Publisher: Elsevier BV

Date: 03-2019

DOI: 10.1016/J.EBIOM.2019.02.035

Use of technology in endodontics by undergraduate dental students in a south‐eastern state of Brazil

Publisher: Wiley

Date: 09-09-2020

DOI: 10.1111/EJE.12594

Design and Characterization of Inulin Conjugate for Improved Intracellular and Targeted Delivery of Pyrazinoic Acid to Monocytes

Publisher: MDPI AG

Date: 22-05-2019

DOI: 10.3390/PHARMACEUTICS11050243

Abstract: The propensity of monocytes to migrate into sites of mycobacterium tuberculosis (TB) infection and then become infected themselves makes them potential targets for delivery of drugs intracellularly to the tubercle bacilli reservoir. Conventional TB drugs are less effective because of poor intracellular delivery to this bacterial sanctuary. This study highlights the potential of using semicrystalline delta inulin particles that are readily internalised by monocytes for a monocyte-based drug delivery system. Pyrazinoic acid was successfully attached covalently to the delta inulin particles via a labile linker. The formation of new conjugate and amide bond was confirmed using zeta potential, Proton Nuclear Magnetic Resonance (1HNMR) and Fourier transform infrared spectroscopy (FTIR). Scanning electron microscopy (SEM) confirmed that no significant change in size after conjugation which is an important parameter for monocyte targeting. Thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) were used to establish the change in thermal properties. The analysis of in-vitro release demonstrated pH-triggered drug cleavage off the delta inulin particles that followed a first-order kinetic process. The efficient targeting ability of the conjugate for RAW 264.7 monocytic cells was supported by cellular uptake studies. Overall, our finding confirmed that semicrystalline delta inulin particles (MPI) can be modified covalently with drugs and such conjugates allow intracellular drug delivery and uptake into monocytes, making this system potentially useful for the treatment of TB.

Ghrelin fights against titanium particle-induced inflammatory osteolysis through activation of ss-catenin signaling pathway

Publisher: Springer Science and Business Media LLC

Date: 04-06-2019

DOI: 10.1007/S10753-019-01026-W

Abstract: Aseptic loosening is a major complication of prosthetic joint surgery, in which exaggerated inflammation and impaired osteoblastogenesis are detected. Ghrelin is a recently discovered neuropeptide that is closely associated with inflammatory conditions and bone regeneration. Here, we report that titanium particles inhibited ghrelin expression in MC3T3-E1 cells. Furthermore, exogenous ghrelin effectively inhibited titanium particle-induced inflammation in vitro by interacting with its receptor GHSR1a as an inhibitor of GHSR1a, Dlys repressed the function of ghrelin. Moreover, ghrelin attenuated the impairment of osteoblastogenesis and the exaggeration of osteolysis induced by titanium particles. Furthermore, the protective role of ghrelin in aseptic loosening might be associated with the Wnt/β-catenin signaling pathway. Collectively, these findings suggest that ghrelin might be a potential therapeutic target for wear-debris-induced inflammation and osteolysis.

CoVaccine HT™ adjuvant is superior to Freund's adjuvants in eliciting antibodies against the endogenous alarmin HMGB1

Publisher: Elsevier BV

Date: 12-2016

DOI: 10.1016/J.JIM.2016.09.008

Abstract: Adjuvants are used to enhance the immune response against specific antigens for the production of antibodies, with the choice of adjuvant most critical for poorly immunogenic and self-antigens. This study quantitatively and qualitatively evaluated CoVaccine HT™ and Freund's adjuvants for eliciting therapeutic ovine polyclonal antibodies targeting the endogenous alarmin, high mobility group box-1 (HMGB1). Sheep were immunised with HMGB1 protein in CoVaccine HT™ or Freund's adjuvants, with injection site reactions and antibody titres periodically assessed. The binding affinity of antibodies for HMGB1 and their neutralisation activity was determined in-vitro, with in vivo activity confirmed using a murine model of endotoxemia. Results indicated that CoVaccine HT™ elicited significantly higher antibody tires with stronger affinity and more functional potency than antibodies induced with Freund's adjuvants. These studies provide evidence that CoVaccine HT™ is superior to Freund's adjuvants for the production of antibodies to antigens with low immunogenicity and supports the use of this alternative adjuvant for clinical and experimental use antibodies.

Water on Tap

Publisher: Cambridge University Press

Date: 21-04-2011

DOI: 10.1017/CBO9780511974823

Abstract: In the 1990s and mid-2000s, turbulent political and social protests surrounded the issue of private sector involvement in providing urban water services in both the developed and developing world. Water on Tap explores ex les of such conflicts in six national settings (France, Bolivia, Chile, Argentina, South Africa and New Zealand), focusing on a central question: how were rights and regulation mobilized to address the demands of redistribution and recognition? Two modes of governance emerged: managed liberalization and participatory democracy, often in hybrid forms that complicated simple oppositions between public and private, commodity and human right. The case studies examine the effects of transnational and domestic regulatory frameworks shaping the provision of urban water services, bilateral investment treaties and the contributions of non-state actors such as transnational corporations, civil society organisations and social movement activists. The conceptual framework developed can be applied to a wide range of transnational governance contexts.

Administration of a commercial immunostimulant preparation, EcoActiva as a feed supplement enhances macrophage respiratory burst and the growth rate of snapper (Pagrus auratus, Sparidae (Bloch and Schneider)) in winter

Publisher: Elsevier BV

Date: 04-2003

DOI: 10.1006/FSIM.2002.0441

Abstract: This study investigated the effects of prolonged administration of a commercial beta-glucan based immunostimulant preparation, EcoActiva, in the form of a feed supplement, on non-specific immune parameters and the growth rate of snapper (Pagrus auratus). Fish held at a temperature representing either summer or winter conditions, were s led periodically and assayed for head kidney macrophage activity via in vitro superoxide production, and classical and alternative complement activity. Fish were also weighed monthly and the growth rate determined. Fish fed on a diet supplemented with EcoActiva and held at the winter temperature had a significant enhancement of macrophage superoxide anion production upon stimulation with phorbol myristate acetate (PMA), and this increased activity was maintained throughout the trial. Macrophage activity in fish fed the supplemented diet and held at the summer temperature was also increased. However, EcoActiva failed to increase either classical or alternate complement activity. Most significantly EcoActiva resulted in an increase in growth rates of the fish held at the winter temperature as compared to the control fish, although no difference was seen between the groups held at the summer temperature. These results suggest that routine incorporation of beta-glucan preparations like EcoActivaduring winter may enhance macrophage function and growth rates at a time of increased disease susceptibility and little or no growth.

A screen of mammalian antibodies on snapper ( Pagrus auratus , Sparidae) peripheral blood leukocytes reveals cross reactivity of an anti-human CD3 antibody with a population of mIg − cells

Publisher: Elsevier BV

Date: 09-2001

DOI: 10.1016/S0145-305X(01)00024-6

Abstract: Detailed immunological studies of the teleosts have been h ered by a lack of antibodies against cell-specific markers. Furthermore, where antibodies have been raised, in many instances they have been found to be species-specific. In comparison, many monoclonal and polyclonal antibodies exist with specificities for mammalian proteins and glycoproteins that effectively differentiate leukocyte sub-populations. In this study, we have tested a panel of 54 commercial antibodies against human and murine cell surface receptors for their ability to bind leukocytes isolated from the peripheral blood of snapper (Pagrus auratus). From this panel, one antibody, A452, which is specific for the intracytoplasmic tail of the epsilon (epsilon) chain of the T cell receptor-associated CD3 complex (CD3epsilon) bound to a subpopulation of peripheral blood leukocytes. Mutually exclusive counterstaining was observed when this antibody was used in conjunction with a monoclonal anti-snapper immunoglobulin antibody. This suggests that A452 may be binding to putative snapper T cells.

Solid-state capture and real-time analysis of individual T cell activation via self-assembly of binding multimeric proteins on functionalized materials surfaces

Publisher: Elsevier BV

Date: 2012

DOI: 10.1016/J.ACTBIO.2011.09.001

Abstract: Polyfunctional T cell responses are increasingly underpinning new and improved vaccination regimens. Studies of the nature and extent of these T cell responses may be facilitated if specific T cell populations can be assessed from mixed populations by ligand-mediated capture in a solid-state assay format. Accordingly, we report here the development of a novel strategy for the solid-state capture and real-time activation analyses of in idual cognate T cells which utilizes a spontaneous self-assembly process for generating multimers of biotinylated class I major histocompatibility-peptide complex (MHCp) directly on the solid-state assay surface while also ensuring stability by covalent interfacial binding. The capture surface was constructed by the fabrication of multilayer coatings onto standard slides. The first layer was a thin polymer coating with surface aldehyde groups, onto which streptavidin was covalently immobilized, followed by the docking of multimers of biotinylated MHCp or biotinylated anti-CD45.1 monoclonal antibody. The high binding strength at each step of this immobilization sequence aims to ensure that artefacts such as (partial) detachment, or displacement by proteins from solution, would not interfere with the intended biological assays. The multilayer coating steps were monitored by X-ray photoelectron spectroscopy data indicated that the MHCp proteins self-assembled in a multimeric form onto the streptavidin surface. Immobilized multimeric MHCp demonstrated the capacity to bind and retain antigen-specific T cells from mixed populations of cells onto the solid carrier. Furthermore, real-time confocal microscopic detection and quantification of subsequent calcium flux using paired fluorescent ratiometric probes facilitated the analysis of in idual T cell response profiles, as well as population analyses using a combination of in idual T cell events.

GM-CSF regulates uterine dendritic cell and macrophage maturation and antigen presentation

Publisher: Elsevier BV

Date: 08-2011

DOI: 10.1016/J.JRI.2011.06.009

Dysregulated innate immune function in the aetiopathogenesis of idiopathic inflammatory myopathies

Publisher: Elsevier BV

Date: 2017

DOI: 10.1016/J.AUTREV.2016.09.019

Abstract: The idiopathic inflammatory myopathies (IIMs) are a heterogeneous group of systemic muscle conditions that are believed to be autoimmune in nature. They have distinct pathological features, but the aetiopathogenesis of each subtype remains largely unknown. Recently, there has been increased interest in the complex role the innate immune system plays in initiating and perpetuating these conditions, and how this may differ between subtypes. This article summarises the traditional paradigms of IIM pathogenesis and reviews the accumulating evidence for disturbances in innate immune processes in these rare, but debilitating chronic conditions.

OVERLAPPING T-CELL EPITOPES IN THE GROUP-I ALLERGEN OF DERMATOPHAGOIDES SPECIES RESTRICTED BY HLA-DP AND HLA-DR CLASS-II MOLECULES

Publisher: Elsevier BV

Date: 05-1994

DOI: 10.1016/0091-6749(94)90383-2

Abstract: The induction of IgE antibodies reactive with the group I allergen of Dermatophagoides species (house dust mite [HDM]), which comprise a major component of the allergic immune response in HDM-atopic in iduals, is dependent on the functional activity of specific CD4+ T cells. In this report we demonstrate that for a particular HDM-atopic in idual the T-cell response to the group I allergen of Dermatophagoides pteronyssinus (Der p I) is limited to a single region (residues 101-143) of the protein. By mapping the fine antigen specificity with T-cell clones, we observed that the sequence 101-131 of Der p I contains a cluster of at least three overlapping T-cell epitopes. Analysis of the HLA class II restriction specificity of the T-cell clones revealed that the T-cell epitope, residues 110-131, was restricted by HLA-DRB1*0101. In contrast, peptide Der p I, 110-119 was recognized in association with HLA-DPB1*0402. However, the ability of cloned T cells to proliferate to the peptide Der p I, 107-119 presented by HLA-DPB1*0401, HLA-DPB1*0402, and HLA-DPB1*0501 expressing accessory cells illustrates the heterogeneity of the restriction specificity of this region of Der p I. The application of this information in the design of peptide-based immunotherapy in the management of allergic responses to HDM is discussed.

Type I interferons mediate the innate cytokine response to recombinant fowlpox virus but not the induction of plasmacytoid dendritic cell-dependent adaptive immunity

Publisher: American Society for Microbiology

Date: 07-2010

DOI: 10.1128/JVI.02618-09

Abstract: Type I interferons (IFNs) are considered to be important mediators of innate immunity due to their inherent antiviral activity, ability to drive the transcription of a number of genes involved in viral clearance, and their role in the initiation of innate and adaptive immune responses. Due to the central role of type I IFNs, we sought to determine their importance in the generation of immunity to a recombinant vaccine vector fowlpox virus (FPV). In analyzing the role of type I IFNs in immunity to FPV, we show that they are critical to the secretion of a number of innate and proinflammatory cytokines, including type I IFNs themselves as well as interleukin-12 (IL-12), tumor necrosis factor-alpha (TNF-α), IL-6, and IL-1β, and that deficiency leads to enhanced virus-mediated antigen expression. Interestingly, however, type I IFNs were not required for adaptive immune responses to recombinant FPV even though plasmacytoid dendritic cells (pDCs), the primary producers of type I IFNs, have been shown to be requisite for this to occur. Furthermore, we provide evidence that the importance of pDCs may lie in their ability to capture and present virally derived antigen to T cells rather than in their capacity as professional type I IFN-producing cells.

HPV E7-mediated NCAPH ectopic expression regulates the carcinogenesis of cervical carcinoma via PI3K/AKT/SGK pathway

Publisher: Springer Science and Business Media LLC

Date: 11-12-2020

DOI: 10.1038/S41419-020-03244-9

Abstract: Cervical cancer is one of the most common gynecological tumors in the world, and human papillomavirus (HPV) infection is its causative agent. However, the molecular mechanisms involved in the carcinogenesis of cervical cancer still require clarification. Here we found that knockdown of Non-SMC (Structural Maintenance of Chromosomes) condensin I complex subunit H (NCAPH) gene expression significantly inhibited the proliferation, migration, invasion and epithelial–mesenchymal transition (EMT) of cervical cancer cells in vitro, and restrained xenograft tumor formation in vivo. Intriguingly, HPV E7 could form a positive feedback loop with NCAPH. E7 upregulated NCAPH gene expression via E2F1 which initiated NCAPH transcription by binding to its promoter directly. Silencing of NCAPH reduced E7 transcription via promoting the transition of AP-1 heterodimer from c-Fos/c-Jun to Fra-1/c-Jun. Moreover, the E7-mediated NCAPH overexpression was involved in the activation of the PI3K/AKT/SGK signaling pathway. In vivo, NCAPH expression in cervical cancer tissues was significantly higher than which in normal cervix and high-grade squamous intraepithelial lesion (HSIL) tissues, and its expression was significantly correlated with tumor size, depth of invasion and lymph node metastasis. Patients with high NCAPH expression had a significantly better survival outcomes than those with low-expression, suggesting that NCAPH-induced cell proliferation might sensitize cancer cells to adjuvant therapy. In conclusion, our results revealed the role of NCAPH in the carcinogenesis of cervical cancer in vitro and in vivo. The interaction between E7 and NCAPH expands the mechanism of HPV induced tumorigenesis and that of host genes regulating HPV E7.

A T cell clone with three potential TCR alpha chain rearrangements expresses only one receptor combination at the cell surface.

Publisher: Elsevier BV

Date: 10-1996

DOI: 10.1016/S0161-5890(96)00076-4

Abstract: In determining the T cell receptor (TcR) usage of various T cell clones that recognize peptide antigens derived from allergens, a particular clone (AC20) was found, that apparently expressed three different species of mRNA encoding alpha chains. The logical conclusion that the cells were not clonal was refuted by the finding of only a single beta chain rearrangement. One of the alpha chains (V alpha20), was not in frame, but two V alpha8 transcripts of different lengths were both potentially translatable. Sequence analysis suggested that the shorter transcript was generated by a secondary splice event from the longer, through the use of a splice donor sequence encoded by the J alpha38 gene segment. The efficiency of excision of the intervening sequence is such that approximately equal amounts of the long and short transcripts occur in the steady state pool of mRNA. This phenomenon has been reported previously in TcR alpha rearrangements, but it has never been made clear whether these truncated chains can form a functional TcR. Reconstitution of a TcR negative cell line with these transcripts showed that only the full length alpha chain was able to pair efficiently with the beta chain to generate a functional receptor at the cell surface.

The interplay between surface nanotopography and chemistry modulates collagen I and III deposition by human dermal fibroblasts

Publisher: American Chemical Society (ACS)

Date: 09-02-2017

DOI: 10.1021/ACSAMI.6B15932

Abstract: The events within the foreign body response are similar to, but ultimately different than, the wound healing cascade. Collagen production by fibroblasts is known to play a vital role in wound healing and device fibrous encapsulation. However, the influence of surface nanotopography on collagen deposition by these cells has not been reported so far. To address this gap, we have developed model substrata having surface nanotopography of controlled height of 16, 38, and 68 nm and tailored outermost surface chemistry of amines, carboxyl acid, and pure hydrocarbon. Fibroblast adhesion was reduced on nanotopographically modified surfaces compared to the smooth control. Furthermore, amine and acid functionalized surfaces showed increased cell proliferation over hydrophobic hydrocarbon surfaces. Collagen III production increased from day 3 to day 8 and then decreased from day 8 to day 16 on all surfaces, while collagen I deposition increased throughout the duration of 16 days. Our data show that the initial collagen I and III deposition can be modulated by selecting desired combinations of surface nanotopography and chemistry. This study provides useful knowledge that could help in tuning fibrous capsule formation and in turn govern the fate of implantable biomaterial devices.

Aberrant Expression of High Mobility Group Box Protein 1 in the Idiopathic Inflammatory Myopathies

Publisher: Frontiers Media SA

Date: 17-04-2020

DOI: 10.3389/FCELL.2020.00226

The potential role of fowlpox virus in rational vaccine design

Publisher: Informa UK Limited

Date: 08-2006

DOI: 10.1586/14760584.5.4.565

Abstract: The design of optimal vaccines requires detailed knowledge of how protective immune responses are generated in vivo under normal circumstances. This approach to vaccine development, where the immune correlates of protection are defined and vaccines are designed to elicit the same response, is called rational vaccine design. Poxviruses are attractive candidates for inclusion in such design strategies owing to their large genome, which allows for the inclusion of multiple heterologous genes, including those encoding antigens, co-stimulatory molecules and cytokines. Fowlpox virus, the prototypical member of the Avipoxvirus genus, is particularly suitable, as it is also incapable of replicating in mammalian cells. The potential of recombinant fowlpox virus as a safe vaccine vector is being evaluated currently in a number of clinical trials for diseases, including HIV, malaria and various types of cancer. Despite their promise, intricate details regarding how fowlpox virus interacts with the host immune system have not been resolved. In this review, the issues surrounding the use of fowlpox virus as a vaccine vector and possible strategies for enhancing its efficacy are discussed.

NS1 DNA vaccination protects against Zika infection through T cell–mediated immunity in immunocompetent mice

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

Date: 06-12-2019

DOI: 10.1126/SCIADV.AAX2388

Abstract: A novel T cell–based ZIKV vaccine, encoding NS1 protein, confers protection against systemic infection.

Utilising T cell receptor transgenic mice to define mechanisms of maternal T cell tolerance in pregnancy

Publisher: Elsevier BV

Date: 12-2010

DOI: 10.1016/J.JRI.2010.05.007

Abstract: Studies in mice demonstrate that the maternal T cell repertoire is aware of paternal antigens during pregnancy, but in healthy pregnancy reactive T cells do not mediate anti-fetal immunity. Mice expressing transgenic T cell receptors (TCRs) specific for paternal and conceptus antigens are powerful tools for elucidating the events surrounding paternal antigen presentation to the maternal T cell repertoire, the nature of the ensuing T cell response and the factors that skew the response towards immune tolerance to allow survival and development of the conceptus. While results from different transgenic TCR models are not always consistent, there is now sufficient data to allow a consensus interpretation that maternal antigen presenting cells present initially seminal fluid antigens and later placenta-derived antigens to both the CD4+ and CD8+ T cell repertoire. T cell proliferation is generally followed by entry into a state of anergy demonstrated by decreased cytokine production and hyporesponsiveness upon restimulation. Some models also demonstrate downregulation of the TCR and co-stimulatory molecules, clonal deletion of paternal antigen-reactive T cells, or alternatively T cell ignorance of paternal antigens. This review will summarise the range of transgenic TCR studies that have shed light on the events surrounding paternal antigen presentation and the various T cell responses to insemination and pregnancy. The benefits, limitations and caveats of these models, and their impact upon data interpretation, are discussed.

The vaccinia‐based Sementis Copenhagen Vector coronavirus disease 2019 vaccine induces broad and durable cellular and humoral immune responses

Publisher: Wiley

Date: 13-03-2022

DOI: 10.1111/IMCB.12539

Abstract: The ongoing coronavirus disease 2019 (COVID‐19) pandemic perpetuated by severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) variants has highlighted the continued need for broadly protective vaccines that elicit robust and durable protection. Here, the vaccinia virus‐based, replication‐defective Sementis Copenhagen Vector (SCV) was used to develop a first‐generation COVID‐19 vaccine encoding the spike glycoprotein (SCV‐S). Vaccination of mice rapidly induced polyfunctional CD8 T cells with cytotoxic activity and robust type 1 T helper‐biased, spike‐specific antibodies, which are significantly increased following a second vaccination, and contained neutralizing activity against the alpha and beta variants of concern. Longitudinal studies indicated that neutralizing antibody activity was maintained up to 9 months after vaccination in both young and middle‐aged mice, with durable immune memory evident even in the presence of pre‐existing vector immunity. Therefore, SCV‐S vaccination has a positive immunogenicity profile, with potential to expand protection generated by current vaccines in a heterologous boost format and presents a solid basis for second‐generation SCV‐based COVID‐19 vaccine candidates incorporating additional SARS‐CoV‐2 immunogens.

Depletion of high-affinity corticosteroid-binding globulin corresponds to illness severity in sepsis and septic shock; clinical implications

Publisher: Wiley

Date: 08-01-2015

DOI: 10.1111/CEN.12680

Abstract: Corticosteroid-binding globulin (CBG) is cleaved by neutrophil elastase converting the high-affinity (haCBG) conformation of CBG to a low-affinity (laCBG) conformation with a ninefold reduced cortisol-binding affinity. These in vitro data suggest that cortisol release by CBG cleavage results in the targeted delivery of cortisol to areas of inflammation. Our objective was to determine whether CBG cleavage alters circulating levels of haCBG and laCBG in vivo in proportion to sepsis severity. Prospective, observational cohort study in an adult tertiary level Intensive Care Unit in Adelaide, Australia. Thirty-three patients with sepsis or septic shock grouped by illness severity [sepsis, septic shock survivors, septic shock nonsurvivors and other shock]. Plasma levels of haCBG and laCBG were assessed using a recently developed in-house assay in patients. Plasma total and free cortisol levels were also measured. Plasma total CBG and haCBG levels fell significantly, in proportion to disease severity (P < 0·0001 for both). There was a nonsignificant increase in free and total cortisol as illness severity worsened (P = 0·19 and P = 0·39, respectively). Illness severity was better correlated with haCBG levels than either free or total cortisol levels. Increasing illness severity in sepsis and septic shock is associated with markedly reduced circulating haCBG concentrations in vivo. We propose that low levels of haCBG in chronic inflammation may limit the availability of cortisol to inflammatory sites, perpetuating the inflammatory process.

Altered superantigenic ligands demonstrate the quantitative nature of T-cell activation

Publisher: Wiley

Date: 12-2000

DOI: 10.1046/J.1440-1711.2000.00971.X

Abstract: In a recent study, a superantigen mutated in the TCR binding site (staphylococcal enterotoxin B (SEB)delta61Y) was described, which behaved as a partial agonist for a Vbeta17-expressing T-cell clone. Evidence is now presented to demonstrate that there is distinct heterogeneity in the response of primary T cells to this protein. Some Vbeta17 T cells responded to SEBdelta61Y by modulating surface receptor expression consistent with activation, and by proliferating. Other Vbeta17 T cells did not proliferate, nor did they display a receptor expression phenotype consistent with activation. However, when repeatedly exposed to the altered superantigen, some of these non-responders entered cell cycle. This pattern of responses was not recapitulated by providing additional costimulation via CD28, although such treatment did induce some of the 'unresponsive' Vbeta17 T cells to upregulate the IL-2 receptor, indicative of partial activation. It was also found that the heterogeneous pattern could be replicated using very low doses of native SEB. The data are discussed in the context of models of T-cell activation in which differences in TCR ligand affinity and dose determine qualitatively different response phenotypes.

Clonal analysis of the atopic immune response to the group 2 allergen of dermatophagoides spp.: Identification of HLA-DR and -DQ restricted T cell epitopes

Publisher: Oxford University Press (OUP)

Date: 1993

DOI: 10.1093/INTIMM/5.12.1589

Abstract: The group 2 allergens of Dermatophagoides spp. (house dust mite, HDM) are a major immunological target for IgE antibodies in the allergic immune response of HDM atopic in iduals. In this report the heterogeneity of the T cell repertoire reactive with the group 2 allergen of Dermatophagoides pteronyssinus (Der p 2) of a HDM allergic in idual was investigated using overlapping synthetic peptides. By clonal analysis four distinct T cell epitopes were identified, located within residues 16-31, 22-40, 82-100, and 111-129. The importance of these epitopes was confirmed by investigation of the peripheral T cell repertoire, with the polyclonal T cell response to Der p 2 failing to show marked variations in epitope specificity over time. Serological inhibition studies and the use of Epstein-Barr virus transformed B cell lines characterized for their expression of HLA-D region gene products demonstrated that recognition of peptides 16-31 and 111-129 was restricted by HLA-DQ (DQB1*0301), whereas peptide 82-100 is recognized in association with HLA-DR (DRB1*1101). Peptide 22-40 was presented by both HLA-DRB1*1101 and -DQB1*0301 class II molecules. The potential application of these findings lies in the design of peptide-based immunotherapeutics for the management of HDM allergic disease.

Hybrid core/shell microparticles and their use for understanding biological processes

Publisher: Elsevier BV

Date: 11-2015

DOI: 10.1016/J.JCIS.2015.06.040

Abstract: Hybrid micro and nanoparticles have become a topic of intense research in recent years. This is due to the special properties of these materials that open new avenues in advanced applications. Herein, we report a novel method for the generation of hybrid particles utilising plasma polymerization. Poly (methyl methacrylate) (PMMA) beads were first coated with a thin allylamine based plasma polymer layer. Gold nanoparticles of engineered size and surface structure were then attached in a controlled manner to the plasma polymer coated beads. To generate uniform chemistry on the outermost surface and to preserve the nanotopography, we deposited a 5-10 nm thin layer of Acpp. We demonstrated that these particles can be utilized in in vivo models to interrogate important biological phenomena. Specifically, we used them in mice to study the inflammatory and foreign body responses to surface nanotopography. The data strongly indicates that surface nanotopography and chemistry can modulate collagen production and the number of adhering immune cells. The method for generating hybrid particles reported here is solvent free and can open new opportunities in fields such as tissue engineering, drug delivery, biosensors, and regenerative medicine.

EphB and Ephrin-B interactions mediate human mesenchymal stem cell suppression of activated T-cells.

Publisher: Mary Ann Liebert Inc

Date: 15-10-2013

DOI: 10.1089/SCD.2012.0676

Controlled release and bioactivity of the monoclonal antibody rituximab from a porous matrix: a potential in situ therapeutic device

Publisher: Elsevier BV

Date: 09-2014

DOI: 10.1016/J.MATLET.2014.05.110

An immunogenic phenotype in paternal antigen‐specific CD8⁺ T cells at embryo implantation elicits later fetal loss in mice

Publisher: Wiley

Date: 27-06-2020

DOI: 10.1038/ICB.2017.41

Abstract: Central to pregnancy success is a state of T cell tolerance to paternal antigens, which is initiated at conception. The role and regulation of specific phenotypes of CD8

Antimicrobial Polymethacrylates Synthesized as Mimics of Tryptophan-Rich Cationic Peptides

Publisher: American Chemical Society (ACS)

Date: 19-03-2014

DOI: 10.1021/MZ5001527

Abstract: This study describes a facile and high yielding route to two series of polymethacrylates inspired by the naturally occurring, tryptophan-rich cationic antimicrobial polymers. Appropriate optimization of indole content within each gave rise to polymers with high potency against

Anti-immunoglobulin binding and activation of snapper (Pagrus auratus) leucocytes

Publisher: Elsevier BV

Date: 04-2002

DOI: 10.1016/S0145-305X(01)00070-2

Abstract: In order to perform specific immunological assays we have produced and characterised three monoclonal antibodies (MAbs) that bind snapper (Pagrus auratus, Bloch and Schneider) immunoglobulin (Ig). Hybridomas were produced and screened for anti-Ig production using ELISA, Western blot and flow cytometry. All three MAbs (designated 2C5, 4A2 and 1C6) bound specifically to the heavy (H) chain of reduced Ig in Western blot. Furthermore, 1C6 was shown to bind to reduced skin mucus Ig H chain and all three MAbs cross-reacted with the H chain of Atlantic salmon and rainbow trout Ig. In flow cytometric analyses 2C5 and 4A2 bound to B cell populations in the peripheral blood and lymphoid organs. Furthermore, cross-linked 2C5 induced an increase in intracellular protein tyrosine phosphorylation in peripheral blood lymphocytes. Phosphorylated proteins exhibited similar molecular weights to those of mammalian Igalpha and Igbeta and may represent snapper mIg accessory molecule analogues. These data exhibit the potential use of 2C5, 4A2 and 1C6 in both cellular and biochemical analyses of populations of snapper leucocytes.

Functionality of Proteins Bound to Plasma Polymer Surfaces

Publisher: American Chemical Society (ACS)

Date: 19-04-2012

DOI: 10.1021/AM300128N

Abstract: The deposition of a thin film layer by plasma polymerization enables the surface functionalization of a wide range of substrate materials for biointerfacial interactions. Plasma polymers can surface-bind proteins specifically via covalent linkages or nonspecifically through other irreversible adsorption mechanisms key questions are whether covalent chemisorption has indeed occurred, and whether the protein retains functionality. Here the mode of surface binding of streptavidin and the biotin binding functionality of the bound streptavidin layer are studied on plasma polymer (pp) surfaces deposited using propionaldehyde and ethanol that were plasma polymerized at different powers (P) to investigate possible mechanisms for protein binding to a range of different surface chemistries. As expected, with pp surfaces composed principally of aldehyde groups, protein conjugation appears to be specific (chemisorption) allowing the immobilization of streptavidin (SAV) molecules retaining the ability to bind biotinylated probes. To contrast with this, we present the first study of protein adsorption to ethanol pp surfaces prepared at different P. This provides an investigation into retention of the hydroxyl functionality in the pp at low P and its effect on protein adsorption. Adsorption of human serum albumin (HSA) to ethanol pp was similar to that on propionaldehyde pp except at low P (5 W) where hydroxyl group retention and hydration presumably has a role in reducing protein adsorption. Although we observed SAV adsorption to ethanol pp surfaces at all P, interestingly, the protein lost its ability to bind biotinylated probes. Thus we suggest that irreversible, nonspecific adsorption of protein on ethanol pp surfaces results in apparent protein denaturation despite the hydrophilic nature of the ethanol pp surface. We conclude by making inferences between the pp structure as measured by X-ray photoelectron spectroscopy (XPS) and the related protein adsorption mechanisms.

TGF-β signalling and immunity in prostate tumourigenesis

Publisher: Informa UK Limited

Date: 08-01-2010

DOI: 10.1517/14728220903544507

Abstract: The TGF-beta's are pleiotropic cytokines that regulate multiple cellular functions. Their role in the prostate is important for normal prostate development and also in prostate tumourigenesis. The interactions TGF-beta-mediated signalling has with maintaining prostate health, as well as its role in prostate tumourigenesis and prostate tumour immune evasion, with emphasis on how a breakdown in these interactions may influence disease progression. That TGF-beta influences normal prostate growth and differentiation by regulating the balance between epithelial cell proliferation and apoptosis, and involving the androgen receptor pathway. That TGF-beta protects and maintains prostate stem cells and a review of the contrasting role TGF-beta has in prostate tumourigenesis and tumour development, where TGF-beta acts as a tumour suppressor and then switches roles to become a tumour promoter, and creates a local immunosuppressive niche leading to systemic tumour tolerance. TGF-beta signalling in prostate cancer is a valid target for the treatment of this disease however any therapeutic regimen will require an understanding of all aspects of the TGF-beta-signalling nexus, otherwise by the very pleiotrophic nature of TGF-beta, limited clinical benefits may result.

Targeting prostate cancer cells with hybrid elastin-like polypeptide/liposome nanoparticles

Publisher: Informa UK Limited

Date: 2018

DOI: 10.2147/IJN.S152485

Partial T cell activation with an altered superantigenic ligand

Publisher: Wiley

Date: 02-2000

DOI: 10.1046/J.1440-1711.2000.00880.X

Abstract: T cells have the capacity to respond to ligands as full, weak, partial or null agonists, or indeed as antagonists. In the present paper, it is reported that staphylococcal enterotoxin B (SEB) mutated in a T cell receptor (TCR) contact site (SEBDelta61Y) behaves as an altered ligand for a T cell clone (AC20) that expresses the Vbeta17 TCR. The T cells were partially activated by SEBDelta61Y, as shown by TCR down-modulation and up-regulation of the IL-2 receptor. However, these cells did not secrete IL-2, IL-3, IL-4 or IFN-gamma, nor did they proliferate. Analysis of intracellular protein tyrosine phosphorylation after cellular activation provided further evidence that SEBDelta61Y could transduce a signal via the Vbeta17 TCR. The events following receptor ligation were clearly different when the T cells were stimulated with SEB or SEBDelta61Y, manifested as both quantitatively and qualitatively different patterns of phosphorylation of intracellular substrates. In contrast, only quantitative differences were apparent when a transfectant expressing the same alpha/beta TCR was stimulated with the different superantigens. Together, these results provide the first demonstration that altered TCR ligands are not restricted to peptides substituted at secondary TCR contact residues. Rather, an altered superantigenic ligand mutated in the TCR binding site can behave as a partial agonist.

GM-CSF Is an Essential Regulator of T Cell Activation Competence in Uterine Dendritic Cells during Early Pregnancy in Mice

Publisher: The American Association of Immunologists

Date: 12-2010

DOI: 10.4049/JIMMUNOL.1001374

Abstract: Uterine dendritic cells (DCs) are critical for activating the T cell response mediating maternal immune tolerance of the semiallogeneicfetus. GM-CSF (CSF2), a known regulator of DCs, is synthesized by uterine epithelial cells during induction of tolerance in early pregnancy. To investigate the role of GM-CSF in regulating uterine DCs and macrophages, Csf2-null mutant and wild-type mice were evaluated at estrus, and in the periconceptual and peri-implantation periods. Immunohistochemistry showed no effect of GM-CSF deficiency on numbers of uterine CD11c+ cells and F4/80+ macrophages at estrus or on days 0.5 and 3.5 postcoitum, but MHC class II+ and class A scavenger receptor+ cells were fewer. Flow cytometry revealed reduced CD80 and CD86 expression by uterine CD11c+ cells and reduced MHC class II in both CD11c+ and F4/80+ cells from GM-CSF–deficient mice. CD80 and CD86 were induced in Csf2−/− uterine CD11c+ cells by culture with GM-CSF. Substantially reduced ability to activate both CD4+ and CD8+ T cells in vivo was evident after delivery of OVA Ag by mating with Act-mOVA males or transcervical administration of OVA peptides. This study shows that GM-CSF regulates the efficiency with which uterine DCs and macrophages activate T cells, and it is essential for optimal MHC class II- and class I-mediated indirect presentation of reproductive Ags. Insufficient GM-CSF may impair generation of T cell-mediated immune tolerance at the outset of pregnancy and may contribute to the altered DC profile and dysregulated T cell tolerance evident in infertility, miscarriage, and preecl sia.

Isolation and Structural Characterization of Echinocystic Acid Triterpenoid Saponins from the Australian Medicinal and Food Plant Acacia ligulata

Publisher: American Chemical Society (ACS)

Date: 04-10-2017

DOI: 10.1021/ACS.JNATPROD.7B00437

Abstract: The Australian plant Acacia ligulata has a number of traditional food and medicinal uses by Australian Aboriginal people, although no bioactive compounds have previously been isolated from this species. Bioassay-guided fractionation of an ethanolic extract of the mature pods of A. ligulata led to the isolation of the two new echinocystic acid triterpenoid saponins, ligulatasides A (1) and B (2), which differ in the fine structure of their glycan substituents. Their structures were elucidated on the basis of 1D and 2D NMR, GC-MS, LC-MS/MS, and saccharide linkage analysis. These are the first isolated compounds from A. ligulata and the first fully elucidated structures of triterpenoid saponins from Acacia sensu stricto having echinocystic acid reported as the aglycone. Compounds 1 and 2 were evaluated for cytotoxic activity against a human melanoma cancer cell line (SK-MEL28) and a diploid fibroblast cell line (HFF), but showed only weak activity.

Therapeutic targeting of HMGB1 during experimental sepsis modulates the inflammatory cytokine profile to one associated with improved clinical outcomes

Publisher: Springer Science and Business Media LLC

Date: 19-07-2017

DOI: 10.1038/S41598-017-06205-Z

Abstract: Sepsis remains a significant health burden and a major clinical need exists for therapeutics to d en the excessive and uncontrolled immune activation. Nuclear protein high mobility group box protein 1 (HMGB1) is released following cell death and is a late mediator in sepsis pathogenesis. While approaches targeting HMGB1 have demonstrated reduced mortality in pre-clinical models of sepsis, the impact of HMGB1 blockade on the complex septic inflammatory milieu and the development of subsequent immunosuppression remain enigmatic. Analysis of plasma s les obtained from septic shock patients established an association between increased HMGB1 and non-survival, higher APACHE II scores, and increased pro-inflammatory cytokine responses. Pre-clinically, administration of neutralising ovine anti-HMGB1 polyclonal antibodies improved survival in murine endotoxaemia and caecal ligation and puncture-induced sepsis models, and altered early cytokine profiles to one which corresponded to patterns observed in the surviving patient cohort. Additionally, anti-HMGB1 treated murine sepsis survivors were significantly more resistant to secondary bacterial infection and exhibited altered innate immune cell phenotypes and cytokine responses. These findings demonstrate that anti-HMGB1 antibodies alter inflammation in murine sepsis models and reduce sepsis mortality without potentiating immunosuppression.

Graphene quantum dot embedded hydrogel for dissolved iron sensing

Publisher: Wiley

Date: 30-08-2019

DOI: 10.1002/SLCT.201901779

Homodimerization Controls the Fibroblast Growth Factor 9 Subfamily's Receptor Binding and Heparan Sulfate-Dependent Diffusion in the Extracellular Matrix

Publisher: Informa UK Limited

Date: 09-2009

DOI: 10.1128/MCB.01780-08

Activating T regulatory cells for tolerance in early pregnancy - the contribution of seminal fluid

Publisher: Elsevier BV

Date: 12-2009

DOI: 10.1016/J.JRI.2009.08.003

Abstract: A state of active tolerance mediated by T regulatory (Treg) cells must be functional from the time of embryo implantation to prevent the conceptus from maternal immune attack. Male seminal fluid and ovarian steroid hormones are implicated in regulating the size and suppressive function of the Treg cell pool during the peri-implantation phase of early pregnancy. Evidence that antigens and cytokine signals in seminal fluid regulate the maternal immune response includes the following: (1) the Treg cell-inducing cytokine TGFbeta and male alloantigens are present in seminal fluid (2) seminal fluid delivery at coitus is sufficient to induce a state of active immune tolerance to paternal alloantigen, even in the absence of conceptus tissue (3) female dendritic cells can cross-present seminal fluid antigens to activate both CD8(+) and CD4(+) T cells, and (4) mating events deficient in either sperm or seminal plasma result in diminished CD4(+) CD25(+) Foxp3(+) Treg cell populations at the time of embryo implantation. Ongoing studies indicate that the cytokine environment during priming to male seminal fluid antigens influences the phenotype of responding T cells, and impacts fetal survival in later gestation. Collectively, these observations implicate factors in the peri-conceptual environment of both male and female origin as important determinants of maternal immune tolerance. Defining the mechanisms controlling tolerance induction will be helpful for developing new therapies for immune-mediated pathologies of pregnancy such as miscarriage and pre-ecl sia.

PECAM-1-regulated signalling thresholds control tolerance in anergic transgenic B-cells

Publisher: Elsevier BV

Date: 03-2008

DOI: 10.1016/J.MOLIMM.2007.09.024

Abstract: Platelet Endothelial Cell Adhesion Molecule-1 (PECAM-1/CD31) is an immunoglobulin (Ig)-immunoreceptor tyrosine based inhibitory motif (Ig-ITIM) superfamily member that recruits and activates protein-tyrosine phosphatases, predominantly SHP-2 and to a lesser extent, SHP-1. Previously, we have shown that deletion of PECAM-1 results in a hyper-proliferative B-cell phenotype. We wanted to test whether the Ig-ITIM superfamily member, PECAM-1 maintains peripheral tolerance by regulating signalling thresholds of B-cells that control autoantibody production or relaxed negative selection of autoreactive B-cells in bone marrow. In order to address this issue, we utilised the classical model of lysozyme/immunoglobulin transgenic mouse model that defines thresholds for eliminating or inactivating self-reactive B-cells. In this study, we show that breeding of double transgenes: soluble hen egg lysozyme (HEL) and its corresponding high-affinity receptor (HEL-Ig) onto PECAM-1 null background resulted in a spontaneous loss of B-cell tolerance in vivo. The resultant PECAM-1(-/-) Dbl Tg mice displayed elevated levels of anti-HEL immunoglobulin M (IgM) antibodies in the serum compared to PECAM-1+/+ anergic counterparts. Dbl Tg B-cells lacking PECAM-1 showed enhanced B-cell proliferation and calcium flux responses to LPS, IL-4 alone, IgM cross-linking and IL-4 indicating augmentation of antigen-receptor signalling. Thus, PECAM-1 is important in maintaining peripheral tolerance in Dbl Tg B-cells.

Unravelling the complexity of cancer–immune system interplay

Publisher: Informa UK Limited

Date: 06-2010

DOI: 10.1586/ERA.10.66

Abstract: The immune system has an intricate and complex relationship with tumorigenesis while it has the capacity to identify and eliminate cancerous cells, the emergence of a tumor signifies its failure to do this. Thus, the immune-tumor interplay is paradoxical as through initial suppression of tumor growth, an immunologically silent or even suppressive tumor evolves. Furthermore, certain immune processes, such as chronic inflammation and immunosuppression, can facilitate malignant progression. Nevertheless, immunotherapeutic approaches can manipulate the immune milieu to improve the therapeutic outcomes of cancer treatments. Furthermore, particular conventional cancer therapies also have immunostimulatory properties in their own right. An in-depth understanding of the intimate involvement of the immune system in tumorigenesis and the potential to manipulate this interaction to improve disease outcomes will enable the development of new and broadly effective cancer therapies.

Conceptus antigens activate the maternal immune response in pregnancy utilising maternal antigen presenting cells

Publisher: Elsevier BV

Date: 10-2006

DOI: 10.1016/J.JRI.2006.08.022

Surface chemistry mediated albumin adsorption, conformational changes and influence on innate immune responses

Publisher: Elsevier BV

Date: 09-2022

DOI: 10.1016/J.APSUSC.2022.153518

Induction of T cell responses to the invariant chain derived peptide CLIP in mice immunized with the group 1 allergen of house dust mite

Publisher: Oxford University Press (OUP)

Date: 1996

DOI: 10.1093/INTIMM/8.7.1091

Abstract: In this study we demonstrate that immunization of H-2(b) mice with the allergen Der p 1 induces MHC class II restricted T cells that proliferate to residues 15-29 of Der p 1 (p15-29) and to the murine MHC class II-associated invariant chain derived peptide (CLIP). T cells from naive H-2(b) mice and those immunized with murine CLIP fail to respond to either CLIP or p15-29. T cell lines and clones reactive with p15-29 strongly proliferate in response to splenic antigen-presenting cells (APC) from normal H-2(b) mice but show reduced proliferation to APC from invariant chain deficient mice. Furthermore, T cells isolated from Der p 1 primed mice and expanded on H-2(b) spleen cells in the absence of the p15-29 epitope retained specificity for both p15-29 and CLIP, suggesting that naturally presented self components can act as mimetic peptides and may maintain T cell memory to foreign antigens.

GM-CSF regulates uterine macrophage and dendritic cell maturation and antigen presentation

Publisher: Elsevier BV

Date: 08-2010

DOI: 10.1016/J.JRI.2010.06.057

Uterine B Cells Exhibit Regulatory Properties During the Peri-Implantation Stage of Murine Pregnancy

Publisher: Frontiers Media SA

Date: 11-12-2019

DOI: 10.3389/FIMMU.2019.02899

Improving vaccines by incorporating immunological coadjuvants

Publisher: Informa UK Limited

Date: 08-2007

DOI: 10.1586/14760584.6.4.559

Abstract: While vaccination continues to be the most successful interventionist health policy to date, infectious disease remains a significant cause of death worldwide. A primary reason that vaccination is not able to generate effective immunity is a lack of appropriate adjuvants capable of initiating the desired immune response. Adjuvant combinations can potentially overcome this problem however, the possible permutations to consider, which include the route and kinetics of vaccination, as well as combinations of adjuvants, are practically limitless. This review aims to summarize the current understanding of adjuvants and related immunological processes and how this knowledge can and has been applied to the strategic selection of adjuvant combinations as components of vaccines against human infectious disease.

Parent/carers' opinions about COVID‐19 vaccination for children with chronic lung diseases

Publisher: Wiley

Date: 10-2021

DOI: 10.1002/HSR2.410

Current and emerging immunotherapeutic approaches to treat and prevent peanut allergy

Publisher: Informa UK Limited

Date: 12-2012

DOI: 10.1586/ERV.12.119

Abstract: Peanut-allergen hypersensitivity reactions, which can result in anaphylactic episodes and death, affect approximately 1% of the general population. Currently, strict avoidance of allergenic food is the only available treatment for this food-induced allergic reaction however, the innocuous presence of trace amounts of peanut protein contaminating food products makes avoidance extremely difficult, especially in children. Therefore, safe and inexpensive therapeutic strategies aimed at prevention and treatment of peanut allergies is urgently required. This review summarizes the current state of knowledge of adaptive immune recognition and responsiveness to peanut allergens and how this can be integrated and subverted into new therapeutic treatment regimens for these dangerous allergic responses. The potential for new strategic vaccination-based interventions to either moderate or prevent these types of responses from occurring is also discussed.

Trophoblasts promote induction of a regulatory phenotype in B cells that can protect against detrimental T cell–mediated inflammation

Publisher: Wiley

Date: 06-10-2019

DOI: 10.1111/AJI.13187

Antibacterial Plasma Polymer Films Conjugated with Phospholipid Encapsulated Silver Nanoparticles

Publisher: American Chemical Society (ACS)

Date: 09-11-2015

DOI: 10.1021/ACSBIOMATERIALS.5B00338

ANALYSIS OF HUMAN T-CELL RESPONSES TO THE GROUP-II ALLERGEN OF DERMATOPHAGOIDES SPECIES - LOCALIZATION OF MAJOR ANTIGENIC SITES

Publisher: Elsevier BV

Date: 07-1993

DOI: 10.1016/0091-6749(93)90044-G

Abstract: IgE antibodies reactive with the group II allergens of Dermatophagoides species (house dust mite [HDM]) are a major component of the allergic immune response in HDM-allergic atopic in iduals. Here we demonstrate, with the use of overlapping synthetic peptides of the group II allergen of Dermatophagoides pteronyssinus (Der p II), that polyclonal T cells isolated from the majority of atopic HDM-allergic in iduals and healthy nonatopic control subjects respond to Der p II and that T-cell epitopes are present in all regions of the protein. From comparison of peptide-specific T-cell proliferation in both groups of in iduals, it appears that together peptides 61-86 and 78-104 are the most frequently recognized (16 of 18 in iduals). We also observed that nine of the 18 in iduals responded to T-cell epitopes in the region 11-50, and with Der p II-reactive T-cell clones, three distinct T-cell epitopes were mapped within the sequence 11-35. Also, with the use of T-cell clones, two additional epitopes were identified at residues 81-96 and 91-101. These results suggest that T-cell epitopes located in these regions (11-50 and 61-104) are immunodominant. The value of this information in the potential application of Der p II peptides to desensitize HDM allergic responses is discussed.

Immobilized Streptavidin Gradients as Bioconjugation Platforms

Publisher: American Chemical Society (ACS)

Date: 27-01-2012

DOI: 10.1021/LA204714P

Abstract: Surface density gradients of streptavidin (SAV) were created on solid surfaces and demonstrated functionality as a bioconjugation platform. The surface density of immobilized streptavidin steadily increased in one dimension from 0 to 235 ng cm(-2) over a distance of 10 mm. The density of coupled protein was controlled by its immobilization onto a polymer surface bearing a gradient of aldehyde group density, onto which SAV was covalently linked using spontaneous imine bond formation between surface aldehyde functional groups and primary amine groups on the protein. As a control, human serum albumin was immobilized in the same manner. The gradient density of aldehyde groups was created using a method of simultaneous plasma copolymerization of ethanol and propionaldehyde. Control over the surface density of aldehyde groups was achieved by manipulating the flow rates of these vapors while moving a mask across substrates during plasma discharge. Immobilized SAV was able to bind biotinylated probes, indicating that the protein retained its functionality after being immobilized. This plasma polymerization technique conveniently allows virtually any substrate to be equipped with tunable protein gradients and provides a widely applicable method for bioconjugation to study effects arising from controllable surface densities of proteins.

Making an Avipoxvirus Encoding a Tumor-Associated Antigen and a Costimulatory Molecule

Publisher: Springer New York

Date: 2014

DOI: 10.1007/978-1-4939-0345-0_32

Abstract: Fowlpox virus (FPV) is a double-stranded DNA virus with a history of use as a live attenuated vaccine in commercial poultry production systems. FPV is also highly amenable to genetic engineering, with a large cloning capacity and many nonessential sites available for integration, meaning that in recombinant form, several transgenes can be expressed simultaneously. Recombinant FPV has proven an effective prophylactic vaccine vector for other diseases of birds, as well as other animal species (Brun et al., Vaccine 26:6508-6528, 2008). These vectors do not integrate into the host genome nor do they undergo productive replication in mammalian cells thus they have a proven and impeccable safety profile and have been progressed as prophylactic and therapeutic vaccine vectors for use in humans (Beukema et al., Expert Rev Vaccines 5:565-577, 2006 Lousberg et al., Expert Rev Vaccines 10:1435-1449, 2011). Furthermore, repeated immunization with FPV does not blunt subsequent vaccine responses, presumably because it is replication-defective, and thus larger doses can be routinely administered (Brun et al., Vaccine 26:6508-6528, 2008). This strengthens the case for FPV as a viable platform vaccine vector, as it means it can be used repeatedly in an in idual to achieve different immunological outcomes. Here we describe in detail the construction of a recombinant variant of FPV expressing the prostate tumor-associated antigen prostatic acid phosphatase (PAP) in conjunction with the immunostimulatory cytokine, interleukin-2 (IL-2), which, if undertaken under the appropriate regulatory conditions and with approvals in place, would theoretically be amenable to clinical trial applications.

Dasatinib suppresses in vitro natural killer cell cytotoxicity

Publisher: American Society of Hematology

Date: 15-04-2008

DOI: 10.1182/BLOOD-2008-02-138701

Systems vaccinology analysis of a recombinant vaccinia-based vector reveals diverse innate immune signatures at the injection site

Publisher: Cold Spring Harbor Laboratory

Date: 18-08-2020

DOI: 10.1101/2020.08.17.254938

Abstract: Poxvirus systems have been extensively used as vaccine vectors. Herein a systems vaccinology analysis of intramuscular injection sites provides detailed insights into host innate immune responses, as well as expression of vector and recombinant immunogen genes, after vaccination with a new multiplication defective, vaccinia-based vector, Sementis Copenhagen Vector. Chikungunya and Zika virus immunogen mRNA and protein expression was associated with necrosing skeletal muscle cells surrounded by mixed cellular infiltrates. Adjuvant signatures at 12 hours post-vaccination were dominated by TLR3, 4 and 9, STING, MAVS, PKR and the inflammasome. Th1 cytokine signatures were dominated by IFNγ, TNF and IL1β, and chemokine signatures by CCL5 and CXCL12. Multiple signatures associated with dendritic cell stimulation were evident. By day seven, vaccine transcripts were absent, and cell death, neutrophil, macrophage and inflammation annotations had abated. No compelling arthritis signatures were identified. Such innate systems vaccinology approaches should inform refinements in poxvirus-based vector design.

GD2-specific CAR T Cells Undergo Potent Activation and Deletion Following Antigen Encounter but can be Protected from Activation-induced Cell Death by PD-1 Blockade

Publisher: Elsevier BV

Date: 06-2016

DOI: 10.1038/MT.2016.63

The inhibitory co-receptor, PECAM-1 provides a protective effect in suppression of collagen-induced arthritis

Publisher: Springer Science and Business Media LLC

Date: 2005

DOI: 10.1007/S10875-005-0354-7

Abstract: Studies of PECAM-1(-/-) mice have identified that PECAM-1 functions as an inhibitory co-receptor to modulate immunological responsiveness. In this study, we describe the in vivo consequences of PECAM-1 deficiency in mouse models of collagen-induced arthritis (CIA) and K/BxN passive transfer model that resembles many of the features of human rheumatoid arthritis. Immunization of PECAM-1(-/-) C57BL/6 (H-2b) mice with chicken collagen type II induced CIA with an incidence of 82% by day 49, while 33% of wild-type and 100% of DBA/1 mice developed arthritis in a similar time frame. The mean onset of disease for PECAM-1(-/-) C57BL/6 mice was day 32 compared to day 51 for wild-type C57BL/6 mice and day 18 for DBA/1 mice (H-2q susceptible). In terms of disease severity, the mean maximal arthritic index for PECAM-1(-/-) C57BL/6 mice was comparable to DBA/1 mice (8.91 +/- 0.91 vs 11.67 +/- 0.82). This mean maximal index in PECAM-1(-/-) C57BL/6 mice was significantly higher than wild-type C57BL/6 mice (5.00 +/- 0.73). IgG1 and IgG2b antibody responses against CII were elevated in arthritic PECAM-1(-/-) C57BL/6 mice compared to wild-type C57BL/6 mice. Histological examination of arthritic paws of PECAM-1(-/-) C57BL/6 mice revealed inflammatory infiltrates of lymphocytic/monocytic cells and cartilage/bone destruction similar to CIA-induced DBA/1 arthritic paws. In the K/BxN model, the arthritis was not augmented in PECAM-1(-/-) mice compared to wild-type mice. In contrast, in active CIA, PECAM-1(-/-) mice developed severe disease comparable to susceptible DBA/1 mice and profoundly more severe than C57BL/6 mice, where only one third developed a mild/moderate disease. Together these observations suggest that PECAM-1 plays a crucial role in the suppression of development of autoimmune arthritis.

Effect of Surface Chemical Functionalities on Collagen Deposition by Primary Human Dermal Fibroblasts

Publisher: American Chemical Society (ACS)

Date: 16-10-2015

DOI: 10.1021/ACSAMI.5B08249

Abstract: Surface modification has been identified as an important technique that could improve the response of the body to implanted medical devices. Collagen production by fibroblasts is known to play a vital role in wound healing and device fibrous encapsulation. However, how surface chemistry affects collagen I and III deposition by these cells has not been systematically studied. Here, we report how surface chemistry influences the deposition of collagen I and III by primary human dermal fibroblasts. Amine (NH3), carboxyl acid (COOH), and hydrocarbon (CH3) surfaces were generated by plasma deposition. This is a practically relevant tool to deposit a functional coating on any type of substrate material. We show that fibroblasts adhere better and proliferate faster on amine-rich surfaces. In addition, the initial collagen I and III production is greater on this type of coating. These data indicates that surface modification can be a promising route for modulating the rate and level of fibrous encapsulation and may be useful in informing the design of implantable biomedical devices to produce more predictable clinical outcomes.

The immune function of MHC class II molecules mutated in the putative superdimer interface

Publisher: Springer Science and Business Media LLC

Date: 05-2005

DOI: 10.1007/S11010-005-5281-4

Abstract: Analysis of the crystal structure of human class II (HLA-DR1) molecules suggests that the alphabeta heterodimer may be further ordered as a dimer of heterodimers (superdimer), leading to the hypothesis that T cell receptor dimerisation is a mechanism for initiating signaling events preceding T cell activation. The interface between pairs of molecules is stabilised by both salt bridges, polar and hydrophobic interactions. The residues that form the superdimer interface occur in three areas distinct from the antigen-binding groove. They can be defined as follows: region 1, beta-beta contacts in the helix of the beta1 domain region 2, alpha-alpha contacts near the alpha 1/alpha2 domain junction and region 3 alpha-beta contacts in the alpha2/beta2 domains adjacent to the plasma membrane. To determine whether salt bridges and polar interactions formed within these regions are involved in the immune function of the murine MHC class II molecule, I-A(b), appropriate residues in both the alpha and beta chain were identified and mutated to uncharged alanine. Cell lines transfected with different combinations of mutated alpha and beta chains were generated and tested for MHC class II expression, peptide binding capabilities, and ability to present antigenic peptide to an OVA-specific T cell hybridoma. With the exception of two residues in region 2, the substitutions tested did not modulate MHC class II expression, or peptide binding function. When tested for ability to present peptide to an antigen-specific T cell hybridoma, with the exception of mutations in region 2, the substitutions did not appear to abrogate the ability of I-A(b) to stimulate the T cells. These results suggest that mutation of residues in region 2 of the putative superdimer interface have a gross effect on the ability of I-A(b) to be expressed on the cell surface. However, abrogation of salt bridges in region 1 and 3 do not influence I-A(b) cell surface expression, peptide binding or ability to stimulate antigen-specific T cells.

From crescent to mature virion: Vaccinia virus assembly and maturation

Publisher: MDPI AG

Date: 07-10-2014

DOI: 10.3390/V6103787

Antigen-specific T-cell responses to a recombinant fowlpox virus are dependent on MyD88 and interleukin-18 and independent of toll-like receptor 7 (TLR7)- and TLR9-mediated innate immune recognition

Publisher: American Society for Microbiology

Date: 04-2011

DOI: 10.1128/JVI.02000-10

Abstract: Fowlpox virus (FWPV) is a double-stranded DNA virus long used as a live-attenuated vaccine against poultry diseases, but more recent interest has focused on its use as a mammalian vaccine vector. Here, in a mouse model system using FWPV encoding the nominal target antigen chicken ovalbumin (OVA) (FWPV OVA ), we describe for the first time some of the fundamental processes by which FWPV engages both the innate and adaptive immune systems. We show that Toll-like receptor 7 (TLR7) and TLR9 are important for type I interferon secretion by dendritic cells, while TLR9 is solely required for proinflammatory cytokine secretion. Despite this functional role for TLR7 and TLR9 in vitro , only the adapter protein myeloid differentiation primary response gene 88 (MyD88) was shown to be essential for the formation of adaptive immunity to FWPV OVA in vivo . The dependence on MyD88 was confined only to the T-cell compartment and was not related to its contribution to TLR signaling, dendritic cell maturation, or the capture and presentation of FWPV-derived OVA antigen. We demonstrate that this is not by means of mediating T-cell-dependent interleukin-1 (IL-1) signaling, but rather, we suggest that MyD88 functions to support T-cell-specific IL-18 receptor signaling, which in turn is essential for the formation of adaptive immunity to FWPV-encoded OVA.

Nanotopography‐Induced Unfolding of Fibrinogen Modulates Leukocyte Binding and Activation

Publisher: Wiley

Date: 20-01-2019

DOI: 10.1002/ADFM.201807453

The multifunctional alarmin HMGB1 with roles in the pathophysiology of sepsis and cancer

Publisher: Wiley

Date: 25-06-2013

DOI: 10.1038/ICB.2013.25

Abstract: Although originally described as a highly conserved nuclear protein involved in DNA replication, transcription and repair, high-mobility group box-1 protein (HMGB1) has emerged as a key mediator in the regulation of immune responses to infection and sterile injury by exhibiting all the properties of a prototypic 'alarmin'. These include rapid passive release in response to pathogenic infection and/or traumatic injury, active secretion providing for chemotactic and cytokine-like function and an ability to resolve inflammation, including tissue repair and remodelling. In this review, we will give an overview of the post-translational modifications necessary for such ersity in biological activity, concentrating particularly on how differences in oxidation of highly conserved redox-sensitive cysteine residues can potentiate inflammatory responses and dictate cellular fate. We will also review the most recent literature on HMGB1 and its involvement in the pathophysiology of sepsis and cancer, as well as cancer therapy-induced mucositis.

Guanylated Polymethacrylates: A Class of Potent Antimicrobial Polymers with Low Hemolytic Activity

Publisher: American Chemical Society (ACS)

Date: 25-10-2013

DOI: 10.1021/BM401128R

Abstract: We have synthesized a series of copolymers containing both positively charged (amine, guanidine) and hydrophobic side chains ( hiphilic antimicrobial peptide mimics). To investigate the structure-activity relationships of these polymers, low polydispersity polymethacrylates of varying but uniform molecular weight and composition were synthesized, using a reversible addition-fragmentation chain transfer (RAFT) approach. In a facile second reaction, pendant amine groups were converted to guanidines, allowing for direct comparison of cation structure on activity and toxicity. The guanidine copolymers were much more active against Staphylococcus epidermidis and Candida albicans compared to the amine analogues. Activity against Staphylococcus epidermidis in the presence of fetal bovine serum was only maintained for guanidine copolymers. Selectivity for bacterial over mammalian cells was assessed using hemolytic and hemagglutination toxicity assays. Guanidine copolymers of low to moderate molecular weight and hydrophobicity had high antimicrobial activity with low toxicity. Optimum properties appear to be a balance between charge density, hydrophobic character, and polymer chain length. In conclusion, a suite of guanidine copolymers has been identified that represent a new class of antimicrobial polymers with high potency and low toxicity.

Tandem peptide epitopes facilitate CD4-dependent activation of T cell clones

Publisher: Wiley

Date: 04-1997

DOI: 10.1038/ICB.1997.20

Abstract: Peptides that consist of two tandemly repeated epitopes joined by a flexible linker have an increased affinity for class II molecules and are more potent at inducing proliferation of T cell clones than monomeric epitopes. The increase in potency of peptides with two epitopes for in idual T cell clones is proportional to the relative CD4 dependence of the clones. We show that epitope dimers activate T cell clones that respond sub-optimally to monomeric epitopes presented by APC from HIV-infected donors. We hypothesize that HIV+ APC normally fail to stimulate the clones because virally encoded gp 120 sequesters CD4 from the activation complex, but epitope dimers overcome this effect because they are better able to recruit CD4. The alpha beta heterodimer of human class II (HLA-DR1) is further ordered as a dimer of heterodimers (superdimer) at least in its crystal form. Since class II molecules have an open-ended antigen binding groove, the superdimer is theoretically permissive of stable binding of two peptide epitopes linked in tandem. Our data support a role for the MHC class II dimer of heterodimers in lifying the proliferative response of T cells to antigen by dint of the superdimers having a higher affinity for CD4 than the nominal class II alpha beta heterodimers.

Poxvirus-based vector systems and the potential for multi-valent and multi-pathogen vaccines

Publisher: Informa UK Limited

Date: 03-10-2018

DOI: 10.1080/14760584.2018.1522255

Abstract: With the increasing number of vaccines and vaccine-preventable diseases, the pressure to generate multi-valent and multi-pathogen vaccines grows. Combining in idual established vaccines to generate single-shot formulations represents an established path, with significant ensuing public health and cost benefits. Poxvirus-based vector systems have the capacity for large recombinant payloads and have been widely used as platforms for the development of recombinant vaccines encoding multiple antigens, with considerable clinical trials activity and a number of registered and licensed products. Herein we discuss design strategies, production processes, safety issues, regulatory hurdles and clinical trial activities, as well as pertinent new technologies such as systems vaccinology and needle-free delivery. Literature searches used PubMed, Google Scholar and clinical trials registries, with a focus on the recombinant vaccinia-based systems, Modified Vaccinia Ankara and the recently developed Sementis Copenhagen Vector. Vaccinia-based platforms show considerable promise for the development of multi-valent and multi-pathogen vaccines, especially with recent developments in vector technologies and manufacturing processes. New methodologies for defining immune correlates and human challenge models may also facilitate bringing such vaccines to market.

Barx2 and Pax7 Have Antagonistic Functions in Regulation of Wnt Signaling and Satellite Cell Differentiation

Publisher: Oxford University Press (OUP)

Date: 23-05-2014

DOI: 10.1002/STEM.1674

Abstract: The canonical Wnt signaling pathway is critical for myogenesis and can induce muscle progenitors to switch from proliferation to differentiation how Wnt signals integrate with muscle-specific regulatory factors in this process is poorly understood. We previously demonstrated that the Barx2 homeobox protein promotes differentiation in cooperation with the muscle regulatory factor (MRF) MyoD. Pax7, another important muscle homeobox factor, represses differentiation. We now identify Barx2, MyoD, and Pax7 as novel components of the Wnt effector complex, providing a new molecular pathway for regulation of muscle progenitor differentiation. Canonical Wnt signaling induces Barx2 expression in muscle progenitors and perturbation of Barx2 leads to misregulation of Wnt target genes. Barx2 activates two endogenous Wnt target promoters as well as the Wnt reporter gene TOPflash, the latter synergistically with MyoD. Moreover, Barx2 interacts with the core Wnt effectors β-catenin and T cell-factor 4 (TCF4), is recruited to TCF/lymphoid enhancer factor sites, and promotes recruitment of β-catenin. In contrast, Pax7 represses the Wnt reporter gene and antagonizes the activating effect of Barx2. Pax7 also binds β-catenin suggesting that Barx2 and Pax7 may compete for interaction with the core Wnt effector complex. Overall, the data show for the first time that Barx2, Pax7, and MRFs can act as direct transcriptional effectors of Wnt signals in myoblasts and that Barx2 and Wnt signaling participate in a regulatory loop. We propose that antagonism between Barx2 and Pax7 in regulation of Wnt signaling may help mediate the switch from myoblast proliferation to differentiation. Stem Cells 2014 :1661–1673

Effect of ‘tongue tie’ use on racing performance of Thoroughbreds in the United Kingdom

Publisher: Wiley

Date: 11-2009

DOI: 10.2746/042516409X434134

Abstract: There is contradictory published evidence on the potential efficacy of 'tongue ties' (TTs) for treatment of intermittent dorsal displacement of the soft palate (DDSP) in racehorses. To evaluate the effect of TTs on racing performance in Thoroughbred racehorses in the U.K. using a retrospective cohort study. Data on in idual horses' lifetime racing performance and TT use were retrieved from the Racing Post Online Database. Exposed cases were horses that ran with a TT in randomly chosen race meetings on one of 60 randomly chosen dates from 2001-2003. Unexposed (control) horses were matched to each exposed horse. Various measures of racing performance were analysed both within and between exposed and unexposed groups. Subsets of exposed horses that ran for 3 or 5 consecutive starts wearing TTs and their matched controls were analysed separately to examine the effect of repeated TT use. The inclusion criteria were fulfilled by 108 horses. The odds ratio for 'improvement' in race earnings between exposed and unexposed horses was 1.85 for horses that ran at least once with a TT, and 3.60 and 4.24, respectively, for horses that ran in 3 or 5 consecutive races wearing a TT. After instigation of TT use, horses that ran in 3 or 5 consecutive races wearing a TT had a significant increase in earnings when they ran wearing a TT compared to their pre-TT races. The use of a TT appears to have a beneficial effect on racing performance in a selected population of Thoroughbred racehorses.

Potent and orally bioavailable CDK8 inhibitors: design, synthesis, structure-activity relationship analysis and biological evaluation

Publisher: Elsevier BV

Date: 03-2021

DOI: 10.1016/J.EJMECH.2021.113248

Snapper (Pagrus auratus) leucocyte proliferation is synergistically enhanced by simultaneous stimulation with LPS and PHA

Publisher: Elsevier BV

Date: 03-2004

DOI: 10.1016/S1050-4648(03)00112-8

Barx2 Is Expressed in Satellite Cells and Is Required for Normal Muscle Growth and Regeneration

Publisher: Oxford University Press (OUP)

Date: 18-01-2012

DOI: 10.1002/STEM.777

Abstract: Muscle growth and regeneration are regulated through a series of spatiotemporally dependent signaling and transcriptional cascades. Although the transcriptional program controlling myogenesis has been extensively investigated, the full repertoire of transcriptional regulators involved in this process is far from defined. Various homeodomain transcription factors have been shown to play important roles in both muscle development and muscle satellite cell-dependent repair. Here, we show that the homeodomain factor Barx2 is a new marker for embryonic and adult myoblasts and is required for normal postnatal muscle growth and repair. Barx2 is coexpressed with Pax7, which is the canonical marker of satellite cells, and is upregulated in satellite cells after muscle injury. Mice lacking the Barx2 gene show reduced postnatal muscle growth, muscle atrophy, and defective muscle repair. Moreover, loss of Barx2 delays the expression of genes that control proliferation and differentiation in regenerating muscle. Consistent with the in vivo observations, satellite cell-derived myoblasts cultured from Barx2−/− mice show decreased proliferation and ability to differentiate relative to those from wild-type or Barx2+/− mice. Barx2−/− myoblasts show reduced expression of the differentiation-associated factor myogenin as well as cell adhesion and matrix molecules. Finally, we find that mice lacking both Barx2 and dystrophin gene expression have severe early onset myopathy. Together, these data indicate that Barx2 is an important regulator of muscle growth and repair that acts via the control of satellite cell proliferation and differentiation. Disclosure of potential conflicts of interest is found at the end of this article.

Plasma polymerised polyoxazoline thin films for biomedical applications

Publisher: Royal Society of Chemistry (RSC)

Date: 2015

DOI: 10.1039/C5CC00260E

Abstract: We report novel solvent-free and substrate independent, plasma polymerised nanoscale biocompatible polyoxazoline coatings capable of controlling protein and cell adhesion, and significantly reducing biofilm build up.

Innate Immunity and Biomaterials at the Nexus: Friends or Foes

Publisher: Hindawi Limited

Date: 2015

DOI: 10.1155/2015/342304

Abstract: Biomaterial implants are an established part of medical practice, encompassing a broad range of devices that widely differ in function and structural composition. However, one common property amongst biomaterials is the induction of the foreign body response: an acute sterile inflammatory reaction which overlaps with tissue vascularisation and remodelling and ultimately fibrotic encapsulation of the biomaterial to prevent further interaction with host tissue. Severity and clinical manifestation of the biomaterial-induced foreign body response are different for each biomaterial, with cases of incompatibility often associated with loss of function. However, unravelling the mechanisms that progress to the formation of the fibrotic capsule highlights the tightly intertwined nature of immunological responses to a seemingly noncanonical “antigen.” In this review, we detail the pathways associated with the foreign body response and describe possible mechanisms of immune involvement that can be targeted. We also discuss methods of modulating the immune response by altering the physiochemical surface properties of the biomaterial prior to implantation. Developments in these areas are reliant on reproducible and effective animal models and may allow a “combined” immunomodulatory approach of adapting surface properties of biomaterials, as well as treating key immune pathways to ultimately reduce the negative consequences of biomaterial implantation.

A vaccinia-based single vector construct multi-pathogen vaccine protects against both Zika and chikungunya viruses

Publisher: Springer Science and Business Media LLC

Date: 26-03-2018

DOI: 10.1038/S41467-018-03662-6

Abstract: Zika and chikungunya viruses have caused major epidemics and are transmitted by Aedes aegypti and/or Aedes albopictu s mosquitoes. The “Sementis Copenhagen Vector” (SCV) system is a recently developed vaccinia-based, multiplication-defective, vaccine vector technology that allows manufacture in modified CHO cells. Herein we describe a single-vector construct SCV vaccine that encodes the structural polyprotein cassettes of both Zika and chikungunya viruses from different loci. A single vaccination of mice induces neutralizing antibodies to both viruses in wild-type and IFNAR −/− mice and protects against (i) chikungunya virus viremia and arthritis in wild-type mice, (ii) Zika virus viremia and fetal lacental infection in female IFNAR −/− mice, and (iii) Zika virus viremia and testes infection and pathology in male IFNAR −/− mice. To our knowledge this represents the first single-vector construct, multi-pathogen vaccine encoding large polyproteins, and offers both simplified manufacturing and formulation, and reduced “shot burden” for these often co-circulating arboviruses.

Polyinosinic: Polycytidylic Acid and Murine Cytomegalovirus Modulate Expression of Murine IL-10 and IL-21 in White Adipose Tissue

Publisher: MDPI AG

Date: 22-05-2020

DOI: 10.3390/V12050569

Abstract: White adipose tissue (WAT) produces interleukin-10 and other immune suppressors in response to pathogen-associated molecular patterns (PAMPs). It also homes a subset of B-cells specialized in the production of IL-10, referred to as regulatory B-cells. We investigated whether viral stimuli, polyinosinic: polycytidylic acid (poly(I:C)) or whole replicative murine cytomegalovirus (MCMV), could stimulate the expression of IL-10 in murine WAT using in vivo and ex vivo approaches. Our results showed that in vivo responses to systemic administration of poly(I:C) resulted in high levels of endogenously-produced IL-10 and IL-21 in WAT. In ex vivo WAT explants, a subset of B-cells increased their endogenous IL-10 expression in response to poly(I:C). Finally, MCMV replication in WAT explants resulted in decreased IL-10 levels, opposite to the effect seen with poly(I:C). Moreover, downregulation of IL-10 correlated with relatively lower number of Bregs. To our knowledge, this is the first report of IL-10 expression by WAT and WAT-associated B-cells in response to viral stimuli.

Dasatinib alters the metastatic phenotype of B16-OVA melanoma in vivo

Publisher: Informa UK Limited

Date: 10-2010

DOI: 10.4161/CBT.10.7.12926

Abstract: The Src/Abl tyrosine kinase inhibitor dasatinib is an approved chronic myeloid leukemia treatment and is under investigation for solid tumor therapy. Members of the Src family of kinases (SFKs) are involved in the process of metastasis and dasatinib inhibits the migration and invasiveness of human melanoma cell lines in vitro. SFKs are also involved in immune function and angiogenesis, which both contribute to As active and passive immunotherapies continue to be investigated in metastatic melanoma, we investigated possible interactions between kinase inhibitors and immunotherapies. A murine syngenic model of metastatic melanoma in which B16F10 cells expressed ovalbumin (B16-OVA) was employed and the active immunotherapy comprised immunization with an OVA-expressing recombinant fowlpox virus (FPVOVA).Dasatinib did not affect B16-OVA viability, proliferation, migration or soft agar colony formation. However, depending on drug dose and schedule, differences in the metastatic behavior of B16-OVA were observed in vivo after dasatinib therapy. At a dose of 5 mg/kg/day given before tumor challenge, dasatinib therapy reduced the number of pulmonary metastases. Conversely, a higher dose (25 mg/kg/day), did not affect the number of pulmonary metastases and increased the number of extra-pulmonary metastases. Finally, immunization of B16-OVA-bearing mice with FPVOVA reduced the number of lung metastases. Prior treatment of these mice with dasatinib 5 mg/kg/day did not affect the incidence of lung metastases. Although the mechanisms by which dasatinib alters the metastatic behavior of B16-OVA cells in vivo remain to be determined, we hypothesize that dasatinib acts via multiple tumor-extrinsic processes that include immune function and neoangiogenesis.

Comparative Analysis of Immune Checkpoint Molecules and Their Potential Role in the Transmissible Tasmanian Devil Facial Tumor Disease

Publisher: Frontiers Media SA

Date: 03-05-2017

DOI: 10.3389/FIMMU.2017.00513

Seminal fluid regulates accumulation of FOXP3 ⁺ regulatory T cells in the preimplantation mouse uterus through expanding the FOXP3 ⁺ cell pool and CCL19-mediated recruitment

Publisher: Oxford University Press (OUP)

Date: 08-2011

DOI: 10.1095/BIOLREPROD.110.088591

Abstract: Regulatory T (Treg) cells facilitate maternal immune tolerance of the semiallogeneic conceptus in early pregnancy, but the origin and regulation of these cells at embryo implantation is unclear. During the preimplantation period, factors in the seminal fluid delivered at coitus cause expansion of a CD4(+)CD25(+) putative Treg cell population in the para-aortic lymph nodes draining the uterus. Using flow cytometry, immunohistochemistry, and real-time quantitative PCR (qPCR) for the signature Treg cell transcription factor FOXP3, we confirmed the identity of the expanded lymph node population as FOXP3(+) Treg cells and showed that this is accompanied by a comparable increase in the uterus of FOXP3(+) Treg cells and expression of Foxp3 mRNA by Day 3.5 postcoitum. Seminal plasma was necessary for uterine Treg cell accumulation, as mating with seminal vesicle-deficient males failed to elicit an increase in uterine Treg cells. Furthermore seminal fluid induced expression of mRNA encoding the Treg chemokine CCL19 (MIP3beta), which acts through the CCR7 receptor to regulate Treg cell recruitment and retention in peripheral tissues. Glandular and luminal epithelial cells were identified as the major cellular origins of uterine CCL19, and exposure to both seminal plasma and sperm was required for maximum expression. Together, these results indicate that Treg cells accumulate in the uterus prior to embryo implantation and that seminal fluid is a key regulator of the uterine Treg cell population, operating by both increasing the pool of available Treg cells and promoting their CCL19-mediated recruitment from the circulation into the implantation site.

A rapid technique to determine performance and efficiency of activated carbon water filters

Publisher: IWA Publishing

Date: 19-06-2018

DOI: 10.2166/WS.2017.123

Abstract: The performance of activated carbon water filters, with respect to the breakthrough of dissolved organic matter (DOM) and dangerous trihalomethanes (THMs) from supplied water, has been analysed by fluorescence spectroscopy. Fluorescence spectroscopy has been demonstrated as a viable technique to monitor carbon filter performance, using the fluorescently active DOM species as an indicator. Due to the relationship between DOM and THMs, where DOM is the precursor for THM formation during the chlorine treatment of water, fluorescence spectroscopy can be used to predict the breakthrough of both species from activated carbon filters. In order to establish a versatile measurement technique, the most appropriate fluorescence excitation and emission wavelengths for detecting the DOM in water were firstly determined. These fluorescence measurement parameters were then applied to effluent water s les from carbon filters, over a total filtrate volume of 4,200 L. The total THM concentration in filtered water s les was determined by headspace gas chromatography (HSGC), with the fluorescence and HSGC results showing a high degree of correlation for the amount of DOM and THM respectively. Importantly, this correlation is observed for both of the determined fluorescence measurement parameters, highlighting the validity and versatility of this technique.

Multi-parameter flow cytometric analysis of uterine immune cell fluctuations over the murine estrous cycle

Publisher: Elsevier BV

Date: 02-2016

DOI: 10.1016/J.JRI.2015.11.005

Abstract: Investigating immune cell populations within various reproductive tissues commonly utilises flow cytometric methods. With advances in fluorophore technology and equipment capabilities, multiple cell types from a single tissue s le can be identified by using different combinations of cell surface markers to distinguish specific cell populations. Here a protocol optimized for mouse uterine tissue was used to show the proportional changes in dendritic cells, monocyte/macrophages, T and B cells, NK and NK T cells, and the granulocytes, neutrophils and eosinophils at each of the four stages of the estrous cycle. Importantly, we demonstrate that use of anti-SiglecF or assessment of FSC/SSC plots could be used to differentiate monocyte/macrophage and eosinophil populations that otherwise cannot be distinguished by use of the common combination of antibodies against F4/80 and CD11b. Our results clearly indicate that within the uterus a dynamic population of immune cells resides, with many cell types reaching peak abundance at estrus and metestrus phases of the cycle, consistent with their importance in the response to paternal antigens and/or pathogens encountered after insemination.

Differentially expressed miRNAs in bone after methotrexate treatment

Publisher: Wiley

Date: 12-09-2022

DOI: 10.1002/JCP.30583

Abstract: Previous studies have shown that administration of antimetabolite methotrexate (MTX) caused a reduced trabecular bone volume and increased marrow adiposity (bone/fat switch), for which the underlying molecular mechanisms and recovery potential are unclear. Altered expression of microRNAs (miRNAs) has been shown to be associated with dysregulation of osteogenic and/or adipogenic differentiation by disrupting target gene expression. First, the current study confirmed the bone/fat switch following MTX treatment in precursor cell culture models in vitro. Then, using a rat intensive 5‐once daily MTX treatment model, this study aimed to identify miRNAs associated with bone damage and recovery (in a time course over Days 3, 6, 9, and 14 after the first MTX treatment). RNA isolated from bone s les of treated and control rats were subjected to miRNA array and reverse transcription‐polymerase chain reaction validation, which identified five upregulated miRNA candidates, namely, miR‐155‐5p, miR‐154‐5p, miR‐344g, miR‐6215, and miR‐6315. Target genes of these miRNAs were predicted using TargetScan and miRDB. Then, the protein‐protein network was established via STRING database, after which the miRNA‐key messenger RNA (mRNA) network was constructed by Cytoscape. Functional annotation and pathway enrichment analyses for miR‐6315 were performed by DAVID database. We found that TGF‐β signaling was the most significantly enriched pathway and subsequent dual‐luciferase assays suggested that Smad2 was the direct target of miR‐6315. Our current study showed that miR‐6315 might be a vital regulator involved in bone and marrow fat formation. Also, this study constructed a comprehensive miRNA–mRNA regulatory network, which may contribute to the pathogenesis rognosis of MTX‐associated bone loss and bone marrow adiposity.

Transforming growth factor-beta-mediated signaling in T lymphocytes impacts on prostate-specific immunity and early prostate tumor progression

Publisher: Elsevier BV

Date: 02-2009

DOI: 10.1038/LABINVEST.2008.123

Abstract: T cells are in general tolerant of prostate-specific tumor antigens. That prostate tumor tissue makes transforming growth factor-beta (TGFbeta) is thought to play a role in the induction of T-cell tolerance within the host and to contribute to tumor progression itself. Here we sought to investigate the influence of TGFbeta signaling on prostate antigen-specific T-cell responses as well as prostate tumorogenesis in an autochthonous murine model of the disease. The response of naive and activated ovalbumin (OVA) antigen-specific T cells, which had been rendered incapable of responding to TGFbeta through T-cell-specific transgenic expression of a dominant-negative variant of the TGFbeta receptor II (dnTGFRII), was analyzed after adoptive transfer into prostate OVA-expressing transgenic (POET) mice. The role of TGFbeta signaling in endogenous T cells in mice, which spontaneously form tumors, was also assessed by monitoring prostate tumor formation and progression in F1 progeny of productive matings between transgenic adenocarcinoma of the mouse prostate (TRAMP) and dnTGFRII mice. TGFbeta-resistant CD8(+) T cells proliferated more and produced IFNgamma more readily after OVA stimulation in vitro. OVA-specific T cells did not damage the prostate gland of POET mice irrespective of TGFbeta responsiveness. However, ex vivo activation facilitated entry of TGFbeta-insensitive T cells into the prostate and was associated with prostate tissue damage. Early tumor progression was delayed in TRAMP mice that carried endogenous TGFbeta-insensitive T cells. Together, these results suggest that TGFbeta-signaling represses CD8(+) T-cell responses to a prostate-specific antigen. TGFbeta-mediated repression of T-cell function may include production of IFNgamma, which is known to contribute to tumor immunosurveillance.

The functional contribution of calcium ion flux heterogeneity in T cells

Publisher: Wiley

Date: 31-03-2015

DOI: 10.1038/ICB.2015.34

Abstract: The role of intracellular calcium ion oscillations in T-cell physiology is being increasingly appreciated by studies that describe how unique temporal and spatial calcium ion signatures can control different signalling pathways. Within this review, we provide detailed mechanisms of calcium ion oscillations, and emphasise the pivotal role that calcium signalling plays in directing crucial events pertaining to T-cell functionality. We also describe methods of calcium ion quantification, and take the opportunity to discuss how a deeper understanding of calcium signalling combined with new detection and quantification methodologies can be used to better design immunotherapies targeting T-cell responses.

Rac2-deficient mice display perturbed T-cell distribution and chemotaxis, but only minor abnormalities in TH1 responses

Publisher: Wiley

Date: 06-2002

DOI: 10.1046/J.1440-1711.2002.01077.X

Abstract: The haematopoietic-specific RhoGTPase, Rac2, has been indirectly implicated in T-lymphocyte development and function, and as a pivotal regulator of T Helper 1 (T(H)1) responses. In other haematopoietic cells it regulates cytoskeletal rearrangement downstream of extracellular signals. Here we demonstrate that Rac2 deficiency results in an abnormal distribution of T lymphocytes in vivo and defects in T-lymphocyte migration and filamentous actin generation in response to chemoattractants in vitro. To investigate the requirement for Rac2 in IFN-gamma production and TH1 responses in vivo, Rac2-deficient mice were challenged with Leishmania major and immunized with ovalbumin-expressing cytomegalovirus. Despite a minor skewing towards a T(H)2 phenotype, Rac2-deficient mice displayed no increased susceptibility to L. major infection. Cytotoxic T-lymphocyte responses to cytomegalovirus and ovalbumin were also normal. Although Rac2 is required for normal T-lymphocyte migration, its role in the generation of T(H)1 responses to infection in vivo is largely redundant.

Seminal fluid regulates the uterine Foxp3(+) T regulatory cell population through CCL19-mediated recruitment during the pen-implantation period in mice

Publisher: Elsevier BV

Date: 08-2010

DOI: 10.1016/J.JRI.2010.06.072

Polymer Supported Carbon for Safe and Effective Remediation of PFOA-and PFOS-Contaminated Water

Publisher: American Chemical Society (ACS)

Date: 03-06-2019

DOI: 10.1021/ACSSUSCHEMENG.9B01793

Activated carbon, carbon nanotubes and graphene: materials and composites for advanced water purification

Publisher: MDPI AG

Date: 02-06-2017

DOI: 10.3390/C3020018

PEPTIDE-INDUCED NONRESPONSIVENESS OF HLA-DP RESTRICTED HUMAN T-CELLS REACTIVE WITH DERMATOPHAGOIDES SPP

Publisher: Elsevier BV

Date: 11-1992

DOI: 10.1016/0091-6749(92)90098-M

Abstract: The activation of CD4+ T lymphocytes, which play a central role in allergic inflammation, depends on the recognition of allergen-derived peptides in association with major histocompatibility complex class II gene products. In this report we demonstrate, at a clonal level, that a component of the T-cell repertoire reactive with Dermatophagoides spp. (house dust mite) in atopic in iduals, is restricted by HLA-DP class II molecules. This supports the recent results emerging from genetic epidemiologic studies that indicate positive associations between the HLA-DP phenotype and immune responsiveness to a variety of common allergens. Our findings also reveal that the T cells restricted by HLA-DP recognize a species-specific epitope located in the group I allergen of Dermatophagoides pteronyssinus (residues 101-119). Furthermore, we report that the pretreatment of the T cells restricted by HLA-DP with the Der p I peptide renders them nonresponsive to an immunogenic challenge with house dust mite allergen, and the loss of antigen-dependent proliferation is associated with downregulation of membrane expression of the T-cell antigen receptor. The ability to functionally inactivate T cells restricted by HLA-DP, as well as those that recognize allergen in association with HLA-DR class II molecules, suggests that desensitization with allergen-derived peptides may have therapeutic potential in the management of allergic diseases irrespective of their HLA class II association.

T-cell recognition of a prostate specific antigen is not sufficient to induce prostate tissue destruction

Publisher: Wiley

Date: 05-2006

DOI: 10.1002/PROS.20307

Abstract: The ability of CD8(+) T-cells to induce prostate inflammation was examined using a prostate ovalbumin expressing transgenic mouse (POET) and/or adoptive transfer of T-cell receptor (TCR) transgenic T-cells (OT-I) that specifically recognize ovalbumin. Localization of inflammatory cells to prostate tissue was examined following T-cell activation via endogenous prostatic antigen, recombinant type 5 adenovirus carrying the gene coding ovalbumin (Ad5-mOVA), or adoptive transfer of in vitro antigen stimulated OT-I cells. Ovalbumin specific OT-I cells were activated by autologous prostate antigen and trafficked to the prostate, but did not induce inflammation unless present in overwhelming numbers ( approximately 65% of CD8(+) T-cells). Activation of antigen specific CD8(+) T-cells in vitro (peptide pulsed antigen presenting cells) or in vivo (Ad5-mOVA) induced transitory prostate inflammation, without induction of prostate pathology, regardless of CD4(+) T-cell availability. Inflammation also was observed in OT-I x POET mice but again, pathological effects were not observed. T lymphocytes specific for a prostate antigen are capable of inducing inflammatory infiltration of prostatic tissue rapidly following activation, but do not produce pathological prostate injury.

Synthesis and characterization of pH-sensitive inulin conjugate of isoniazid for monocyte-targeted delivery

Publisher: MDPI AG

Date: 28-10-2019

DOI: 10.3390/PHARMACEUTICS11110555

Abstract: The use of particles for monocyte-mediated delivery could be a more efficient strategy and approach to achieve intracellular targeting and delivery of antitubercular drugs to host macrophages. In this study, the potential of inulin microparticles to serve as a drug vehicle in the treatment of chronic tuberculosis using a monocytes-mediated drug targeting approach was evaluated. Isoniazid (INH) was conjugated to inulin via hydrazone linkage in order to obtain a pH-sensitive inulin-INH conjugate. The conjugate was then characterized using proton nuclear magnetic resonance (1HNMR), Fourier transform infrared spectroscopy (FTIR) as well as in vitro, cellular uptake and intracellular Mycobacterium tuberculosis (Mtb) antibacterial efficacy. The acid-labile hydrazone linkage conferred pH sensitivity to the inulin-INH conjugate with ~95, 77 and 65% of the drug released after 5 h at pH 4.5, 5.2, and 6.0 respectively. Cellular uptake studies confirm that RAW 264.7 monocytic cells efficiently internalized the inulin conjugates into endocytic compartments through endocytosis. The intracellular efficacy studies demonstrate that the inulin conjugates possess a dose-dependent targeting effect against Mtb-infected monocytes. This was through efficient internalization and cleavage of the hydrazone bond by the acidic environment of the lysosome, which subsequently released the isoniazid intracellularly to the Mtb reservoir. These results clearly suggest that inulin conjugates can serve as a pH-sensitive intracellular drug delivery system for TB treatment.

Erratum: Improving vaccines by incorporating immunological coadjuvants (Expert Review Vaccines (2007) 6, 4, (559-578))

Publisher: Informa UK Limited

Date: 12-2007

DOI: 10.1586/14760584.6.6.1022

Innate immune recognition of poxviral vaccine vectors

Publisher: Informa UK Limited

Date: 10-2011

DOI: 10.1586/ERV.11.121

Abstract: The study of poxviruses pioneered the field of vaccinology after Jenner's remarkable discovery that 'vaccination' with the phylogenetically related cowpox virus conferred immunity to the devastating disease of smallpox. The study of poxviruses continues to enrich the field of virology because the global eradication of smallpox provides a unique ex le of the potency of effective immunization. Other poxviruses have since been developed as vaccine vectors for clinical and veterinary applications and include modified vaccinia virus strains such as modified vaccinia Ankara and NYVAC as well as the avipox viruses, fowlpox virus and canarypox virus. Despite the empirical development of poxvirus-based vectored vaccines, it is only now becoming apparent that we need to better understand how the innate arm of the immune system drives adaptive immunity to poxviruses, and how this information is relevant to vaccine design strategies, which are the topics addressed in this article.

Transient dominant host-range selection using Chinese hamster ovary cells to generate marker-free recombinant viral vectors from vaccinia virus

Publisher: Future Science Ltd

Date: 04-2017

DOI: 10.2144/000114537

Abstract: Recombinant vaccinia viruses (rVACVs) are promising antigen-delivery systems for vaccine development that are also useful as research tools. Two common methods for selection during construction of rVACV clones are (i) co-insertion of drug resistance or reporter protein genes, which requires the use of additional selection drugs or detection methods, and (ii) dominant host-range selection. The latter uses VACV variants rendered replication-incompetent in host cell lines by the deletion of host-range genes. Replicative ability is restored by co-insertion of the host-range genes, providing for dominant selection of the recombinant viruses. Here, we describe a new method for the construction of rVACVs using the cowpox CP77 protein and unmodified VACV as the starting material. Our selection system will expand the range of tools available for positive selection of rVACV during vector construction, and it is substantially more high-fidelity than approaches based on selection for drug resistance.

Regulatory B Cells: Dark Horse in Pregnancy Immunotherapy?

Publisher: Elsevier BV

Date: 2021

DOI: 10.1016/J.JMB.2020.07.008

Isolation and partial characterization of a pentraxin-like protein with complement-fixing activity from snapper (Pagrus auratus, Sparidae) serum

Publisher: Elsevier BV

Date: 06-2003

DOI: 10.1016/S0145-305X(03)00034-X

Abstract: Pentraxin-like molecules have been isolated from a number of fish species. However, little is known about the function of these proteins in the teleosts. In this study we report the isolation and characterization of a pentraxin-like molecule from the serum of snapper (Pagrus auratus) that has the ability to activate complement. This pentraxin-like protein was isolated from serum by calcium-dependent binding to agarose. SDS-PAGE analysis demonstrated an oligomeric protein of approximately 200k Da consisting of non-covalently bound subunits of 26 and 23 kDa. Protein sequencing revealed significant (50%) sequence identity with pentraxins from both Atlantic salmon (S. salar) and rainbow trout (O. mykiss). However, polyclonal antibodies raised against snapper pentraxin did not recognise salmon or trout pentraxin in Western blot analysis. Following LPS injection, snapper pentraxin levels increased 2-fold before gradually returning to basal levels. Most significantly, the isolated pentraxin initiated complement-mediated lysis of ligand-coated sheep erythrocytes in a dose-dependent fashion. In view of the similarity between the known fish pentraxins, and their similarity to mammalian serum amyloid P-components we conclude that the isolated protein may be a snapper pentraxin homologue.

Major histocompatibility complex independent clonal T cell anergy by direct interaction of Staphylococcus aureus enterotoxin B with the T cell antigen receptor.

Publisher: Rockefeller University Press

Date: 06-1992

DOI: 10.1084/JEM.175.6.1493

Abstract: The Staphylococcal enterotoxin superantigens stimulate vigorous responses in T cells bearing certain T cell antigen receptor (TCR) V beta regions. In addition to activation, these superantigens also impart negative signals to T cells resulting in a profound state of unresponsiveness or anergy. The Staphylococcus aureus enterotoxins (SE) B and C2 bind to a closely related site on major histocompatibility complex (MHC) human leukocyte antigen (HLA)-DR1 molecules. Only SEB, however, interacts with the TCR V beta 3 region of HA1.7, a human HLA-DR1 restricted T cell clone specific for influenza haemagglutinin. In competition experiments, we demonstrated that the induction of anergy in HA1.7 by SEB is unaffected by the presence of SEC2. These results suggest that SEB-induced anergy is MHC independent and involves a direct interaction between the TCR and SEB. To resolve definitively whether SEB binds directly to T cells in the absence of MHC class II molecules, the cDNAs encoding the HA1.7 TCR were transfected into an MHC class II-negative human T cell line. The addition of SEB to these transfectants resulted in the downregulation of cell surface TCR expression, an increase in the concentration of intracellular calcium ions, the production of lymphokines, and reduced responsiveness to a subsequent challenge with SEB. We conclude that SEB interacts directly with the TCR in the absence of cointeraction with MHC class II molecules, and that this interaction may induce anergy in HA1.7.

Induction of both cellular and humoral immunity following a rational prime-boost immunization regimen that incorporates recombinant ovine atadenovirus and fowlpox virus

Publisher: American Society for Microbiology

Date: 11-2010

DOI: 10.1128/CVI.00291-10

Abstract: Recombinant fowlpox viruses (rFPV) and ovine atadenoviruses (rOAdV) are being developed as safe, nonpathogenic, prophylactic and therapeutic vaccine vectors. There is scope, however, to improve the limited immune responses elicited by each of these vaccine vectors. Using previously determined and optimized routes of administration and viral doses, we characterized the primary adaptive immune responses elicited by recombinant variants of each virus. We demonstrate the contrasting nature of the response elicited by each recombinant virus. Whereas rFPV generates predominately cell-mediated immunity to our nominal target antigen, ovalbumin (OVA), rOAdV drives strong humoral responses. By defining the time taken to achieve maximal cytotoxic T cell responses and by studying the different patterns and kinetics of major histocompatibility complex class I-restricted OVA antigen expression postimmunization, we proposed a heterologous prime-boost regimen of immunization with rOAdV followed by rFPV. The subsequent experimental results showed that this approach produced robust cell-mediated and humoral immune responses against OVA that, importantly, were accompanied by weak anti-viral vector antibody responses. These results, therefore, represent a novel and potentially clinically applicable way to achieve broadly based and effective immunity to the antigens encoded by vectored vaccines.

RAFT-derived antimicrobial polymethacrylates: elucidating the impact of end-groups on activity and cytotoxicity

Publisher: Royal Society of Chemistry (RSC)

Date: 2014

DOI: 10.1039/C4PY00652F

Abstract: We report the use of RAFT polymerization to obtain eight cationic methacrylate polymers bearing amine or guanidine pendant groups, while varying the R- and Z-RAFT end-groups.

Mechanical overloading induces GPX4-regulated chondrocyte ferroptosis in osteoarthritis via Piezo1 channel facilitated calcium influx

Publisher: Elsevier BV

Date: 11-2022

DOI: 10.1016/J.JARE.2022.01.004

The efficacy of a commercial beta-glucan preparation, EcoActiva (TM), on stimulating respiratory burst activity of head-kidney macrophages from pink snapper (Pagrus auratus), Sparidae

Publisher: Elsevier BV

Date: 11-2001

DOI: 10.1006/FSIM.2001.0343

Abstract: This study investigated the in vitro effects of a commercial beta-glucan preparation, EcoActiva, on the respiratory burst activity of head-kidney macrophages isolated from pink snapper (Pagrus auratus), a marine fish cultured in Australia. Macrophages incubated with EcoActiva displayed morphological characteristics of activation, and were stimulated to produce superoxide. Pre-incubation with low levels of EcoActiva significantly increased the response to phorbol myristate acetate (PMA) and lipopolysaccharide (LPS), indicating that EcoActiva could prime these macrophages. Co-culturing macrophages with both LPS and PMA, or EcoActiva and PMA, increased burst activity compared with the response to PMA alone, however, this increase was additive and not synergistic. These results suggest that EcoActiva is able to stimulate non-specific immunity in snapper through increased respiratory burst activity of macrophages, an important component of the host defence network.

Human Flt-3 ligand-mobilized dendritic cells, require additional activation to drive effective immune responses

Publisher: Elsevier BV

Date: 2008

DOI: 10.1016/J.EXPHEM.2007.08.024

Abstract: Dendritic cells (DCs) play a pivotal role in the induction of immunity in response to pathogenic challenge or vaccination. As such, the fms-like tyrosine kinase 3-ligand (Flt-3L) has been used to increase DC populations in vivo, with contrasting outcomes, which include an increase in immunity, tolerance induction, or expansion of regulatory cells. This study examines the adjuvant role that human Flt-3L (hFL) administration has in generating immune responses upon immunization with a poorly immunogenic and soluble protein antigen. Mice were immunized with the nominal antigen, ovalbumin, alone or with antigen emulsified in complete Freund's adjuvant (CFA), with or without prior hFL-mediated expansion of DC subsets. The maturation of DC subsets and activation status of antigen-specific T cells were analyzed by flow cytometry, with effector function assessed in cytolytic T-lymphocyte assays. hFL treatment expanded both conventional DC and plasmacytoid DC in vivo, resulting in increased antigen presentation by both direct and cross-presentation pathways. However, it was only in the context of CFA that antigen immunization could mature DCs and subsequently fully activate antigen-specific T cells with enhanced cytolytic activity. Our studies reveal that hFL essentially acts as a coadjuvant, as hFL augments the size of an immune response but requires further adjuvant activation to alter the quality of the response.

Facial vein injection of human cells in severe combined immunodeficiency (SCID) neonatal mice

Publisher: Elsevier BV

Date: 2018

DOI: 10.1016/J.MEX.2018.10.006

Inflammasome components ASC and AIM2 modulate the acute phase of biomaterial implant-induced foreign body responses

Publisher: Springer Science and Business Media LLC

Date: 10-02-2016

DOI: 10.1038/SREP20635

Abstract: Detailing the inflammatory mechanisms of biomaterial-implant induced foreign body responses (FBR) has implications for revealing targetable pathways that may reduce leukocyte activation and fibrotic encapsulation of the implant. We have adapted a model of poly(methylmethacrylate) (PMMA) bead injection to perform an assessment of the mechanistic role of the ASC-dependent inflammasome in this process. We first demonstrate that ASC −/− mice subjected to PMMA bead injections had reduced cell infiltration and altered collagen deposition, suggesting a role for the inflammasome in the FBR. We next investigated the NLRP3 and AIM2 sensors because of their known contributions in recognising damaged and apoptotic cells. We found that NLRP3 was dispensable for the fibrotic encapsulation however AIM2 expression influenced leukocyte infiltration and controlled collagen deposition, suggesting a previously unexplored link between AIM2 and biomaterial-induced FBR.

The opsonising activity of a pentraxin-like protein isolated from snapper (Pagrus auratus, Sparidae) serum

Publisher: Elsevier BV

Date: 2005

DOI: 10.1016/J.DCI.2004.05.009

Abstract: In mammals the pentraxins, C-reactive protein (CRP) and serum amyloid P component (SAP) are important components of the immune response. Although pentraxins have been isolated from a number of fish species few studies detail their functional immunological role. In this paper we report the establishment of a flow cytometry based assay to measure the phagocytic activity of isolated snapper head kidney leukocytes (HKLs). This assay was then used to examine the ability of a pentraxin-like protein isolated from the serum of snapper (P. auratus) (Sn-PLP) to act as an opsonin. Incubation of snapper head kidney leukocytes (HKL) with FITC-labelled beads resulted in uptake of these particles by approximately 35% of HKLs. Incubation of beads with Sn-PLP enhanced phagocytosis by snapper HKLs in a dose-dependant manner. Enhanced phagocytosis could be inhibited by addition of a rabbit anti-Sn-PLP antibody suggesting that Sn-PLP may act as a ligand for a HKL cell surface receptor. This study provides further evidence toward a functional role for pentraxins in the host defence repertoire of fish.

Nano‐Roughness‐Mediated Macrophage Polarization for Desired Host Immune Response

Publisher: Wiley

Date: 13-08-2023

DOI: 10.1002/SMSC.202300080

Abstract: Macrophage polarization is a significant event in the host immune response, which can be modulated by modifying the surface of a biomaterial. Previous studies have demonstrated the modulation of macrophage polarization using different surface features however, none of these studies reflect the effect of surface properties on unstimulated macrophage polarization for a prolonged period. To better understand the impact of surface features, in this work differentiated THP‐1 cells are employed to control macrophage polarization on nano‐rough surfaces for a duration of 7 days. Model nano‐rough substrates are fabricated by immobilizing gold nanoparticles (AuNPs) of predetermined sizes (16, 38, 68 nm) on a 2‐methyl‐2‐oxazoline thin film, followed by tailoring the outermost surface chemistry. All modified surfaces support high levels of cell adhesion and proliferation. Over time, the expression of pro‐inflammatory cytokines decreases, whereas the expression of anti‐inflammatory cytokines increases on all modified surfaces. Similarly, pro‐inflammatory interleukin (IL)‐1β gene expression is downregulated, and anti‐inflammatory IL‐10‐gene expression is upregulated, regardless of the surface roughness. Analysis of cell morphology reveals that the predominant cell type on the modified surfaces exhibits M2 anti‐inflammatory phenotype. Herein, how surface features can modulate macrophage responses over an extended period is highlighted, offering insights for the development of future biomaterial implants.

Injection site vaccinology of a recombinant vaccinia-based vector reveals diverse innate immune signatures

Publisher: Public Library of Science (PLoS)

Date: 13-01-2021

DOI: 10.1371/JOURNAL.PPAT.1009215

Abstract: Poxvirus systems have been extensively used as vaccine vectors. Herein a RNA-Seq analysis of intramuscular injection sites provided detailed insights into host innate immune responses, as well as expression of vector and recombinant immunogen genes, after vaccination with a new multiplication defective, vaccinia-based vector, Sementis Copenhagen Vector. Chikungunya and Zika virus immunogen mRNA and protein expression was associated with necrosing skeletal muscle cells surrounded by mixed cellular infiltrates. The multiple adjuvant signatures at 12 hours post-vaccination were dominated by TLR3, 4 and 9, STING, MAVS, PKR and the inflammasome. Th1 cytokine signatures were dominated by IFNγ, TNF and IL1β, and chemokine signatures by CCL5 and CXCL12. Multiple signatures associated with dendritic cell stimulation were evident. By day seven, vaccine transcripts were absent, and cell death, neutrophil, macrophage and inflammation annotations had abated. No compelling arthritis signatures were identified. Such injection site vaccinology approaches should inform refinements in poxvirus-based vector design.

The Role of Surface Nanotopography and Chemistry on Primary Neutrophil and Macrophage Cellular Responses

Publisher: Wiley

Date: 04-02-2016

DOI: 10.1002/ADHM.201500845

Abstract: Synthetic materials employed for enhancing, replacing, or restoring biological functionality may be compromised by the host immune responses that they evoke. Surface modification has attracted substantial attention as a tool to modulate the host response to synthetic materials however, how surface nanotopography combined with chemistry affects immune effector cell responses is still poorly understood. To address this open question, a unique set of model surfaces with controlled surface nanotopography in the range of 16, 38, and 68 nm has been generated. Tailored outermost surface chemistry that was amine, carboxyl, or methyl group rich has been provided. The combinations of these properties yield 12 surface types that are subject to functional assays assessing key immune effector cells, namely, primary neutrophil and macrophage responses in vitro. The data demonstrate that surface nanotopography leads to enhanced matrix metalloproteinase-9 production from primary neutrophils, and a decrease in pro-inflammatory cytokine secretion from primary macrophages. Together, these results are the first to directly compare the immunomodulatory effects of the cooperative interplay between surface nanotopography and chemistry.

Individual and Population Quantitative Analyses of Calcium Flux in T-Cells Activated on Functionalized Material Surfaces

Publisher: CSIRO Publishing

Date: 2012

DOI: 10.1071/CH11311

Abstract: We have developed a novel method for activating T-cells on material surfaces that enable in idual and population-based analyses of intracellular calcium flux, as a quantitative measure of T-cell receptor engagement. Functionalized material surfaces were created using a plasma-polymerized foundation layer to immobilize stimulatory T-cell ligands, which could induce T-cell receptor-dependent calcium flux in naive T-cells. Real-time confocal microscopic detection and quantification of calcium flux using paired fluorescent ratiometric probes facilitated the tracking and analysis of response profiles of in idual T-cells, as well as population analyses using a combination of in idual T-cell events. This type of combined analysis cannot be achieved using traditional population-based flow cytometric approaches, and thus provides a logical step towards developing the capacity to assess the magnitude and quality of inherently heterogeneous effector T-cell responses to antigenic challenge.

Self-sterilizing antibacterial silver-loaded microneedles

Publisher: Royal Society of Chemistry (RSC)

Date: 2019

DOI: 10.1039/C8CC06035E

Abstract: Here we report the development of slef-sterilizing dissolving microneedles, a promising vehicle for vaccine and drug delivery.

EphB4 Expressing Stromal Cells Exhibit an Enhanced Capacity for Hematopoietic Stem Cell Maintenance

Publisher: Oxford University Press (OUP)

Date: 23-06-2015

DOI: 10.1002/STEM.2069

Abstract: The tyrosine kinase receptor, EphB4, mediates cross-talk between stromal and hematopoietic populations during bone remodeling, fracture repair and arthritis, through its interactions with the ligand, ephrin-B2. This study demonstrated that transgenic EphB4 mice (EphB4 Tg), over-expressing EphB4 under the control of collagen type-1 promoter, exhibited higher frequencies of osteogenic cells and hematopoietic stem rogenitor cells (HSC), correlating with a higher frequency of long-term culture-initiating cells (LTC-IC), compared with wild type (WT) mice. EphB4 Tg stromal feeder layers displayed a greater capacity to support LTC-IC in vitro, where blocking EphB4/ephrin-B2 interactions decreased LTC-IC output. Similarly, short hairpin RNA-mediated EphB4 knockdown in human bone marrow stromal cells reduced their ability to support high ephrin-B2 expressing CD34+ HSC in LTC-IC cultures. Notably, irradiated EphB4 Tg mouse recipients displayed enhanced bone marrow reconstitution capacity and enhanced homing efficiency of transplanted donor hematopoietic stem rogenitor cells relative to WT controls. Studies examining the expression of hematopoietic supportive factors produced by stromal cells indicated that CXCL12, Angiopoietin-1, IL-6, FLT-3 ligand, and osteopontin expression were more highly expressed in EphB4 Tg stromal cells compared with WT controls. These findings indicate that EphB4 facilitates stromal-mediated support of hematopoiesis, and constitute a novel component of the HSC niche. Stem Cells 2015 :2838—2849

Preserved antiviral adaptive immunity following polyclonal antibody immunotherapy for severe murine influenza infection

Publisher: Springer Science and Business Media LLC

Date: 06-07-2016

DOI: 10.1038/SREP29154

Abstract: Passive immunotherapy may have particular benefits for the treatment of severe influenza infection in at-risk populations, however little is known of the impact of passive immunotherapy on the formation of memory responses to the virus. Ideally, passive immunotherapy should attenuate the severity of infection while still allowing the formation of adaptive responses to confer protection from future exposure. In this study, we sought to determine if administration of influenza-specific ovine polyclonal antibodies could inhibit adaptive immune responses in a murine model of lethal influenza infection. Ovine polyclonal antibodies generated against recombinant PR8 (H1N1) hemagglutinin exhibited potent prophylactic capacity and reduced lethality in an established influenza infection, particularly when administered intranasally. Surviving mice were also protected against reinfection and generated normal antibody and cytotoxic T lymphocyte responses to the virus. The longevity of ovine polyclonal antibodies was explored with a half-life of over two weeks following a single antibody administration. These findings support the development of an ovine passive polyclonal antibody therapy for treatment of severe influenza infection which does not affect the formation of subsequent acquired immunity to the virus.

Bright lights down under: Metal ion complexes turning the spotlight on metabolic processes at the cellular level

Publisher: Elsevier BV

Date: 11-2018

DOI: 10.1016/J.CCR.2017.11.026

The Homeobox Transcription Factor Barx2 Regulates Plasticity of Young Primary Myofibers

Publisher: Public Library of Science (PLoS)

Date: 15-07-2010

DOI: 10.1371/JOURNAL.PONE.0011612

GDF11 antagonizes TNF-a-induced inflammation and protects against the development of inflammatory arthritis in mice

Publisher: Wiley

Date: 08-11-2019

DOI: 10.1096/FJ.201801375RR

Targeting prostate cancer cells with genetically engineered polypeptide-based micelles displaying gastrin-releasing peptide

Publisher: Elsevier BV

Date: 11-2016

DOI: 10.1016/J.IJPHARM.2016.09.039

Abstract: In recent years G protein-coupled receptors (GPCRs) have emerged as crucial tumorigenic factors that drive aberrant cancer growth, metastasis and angiogenesis. Consequently, a number of GPCRs are strongly expressed in cancer derived cell lines and tissue s les. Therefore a rational anti-cancer strategy is the design of nano-medicines that specifically target GPCRs to bind and internalise cytotoxic drugs into cancer cells. Herein, we report the genetic engineering of a self-assembling nanoparticle based on elastin-like polypeptide (ELP), which has been fused with gastrin releasing peptide (GRP). These nanoparticles increased intracellular calcium concentrations when added to GRP receptor positive PC-3 prostate cancer cells, demonstrating specific receptor activation. Moreover, GRP-displaying fluorescent labelled nanoparticles showed specific cell-surface interaction with PC-3 prostate cancer cells and increased endocytic uptake. These nanoparticles therefore provide a targeted molecular carrier system for evaluating the delivery of cytotoxic drugs into cancer cells.

Cortistatin protects against intervertebral disc degeneration through targeting mitochondrial ROS-dependent NLRP3 inflammasome activation

Publisher: Ivyspring International Publisher

Date: 2020

DOI: 10.7150/THNO.45359

The contribution of inflammasome components on macrophage response to surface nanotopography and chemistry

Publisher: Springer Science and Business Media LLC

Date: 18-05-2016

DOI: 10.1038/SREP26207

Abstract: Implantable devices have become an established part of medical practice. However, often a negative inflammatory host response can impede the integration and functionality of the device. In this paper, we interrogate the role of surface nanotopography and chemistry on the potential molecular role of the inflammasome in controlling macrophage responses. To achieve this goal we engineered model substrata having precisely controlled nanotopography of predetermined height and tailored outermost surface chemistry. Bone marrow derived macrophages (BMDM) were harvested from genetically engineered mice deficient in the inflammasome components ASC, NLRP3 and AIM2. These cells were then cultured on these nanoengineered substrata and assessed for their capacity to attach and express pro-inflammatory cytokines. Our data provide evidence that the inflammasome components ASC, NLRP3 and AIM2 play a role in regulating macrophage adhesion and activation in response to surface nanotopography and chemistry. The findings of this paper are important for understanding the inflammatory consequences caused by biomaterials and pave the way to the rational design of future implantable devices having controlled and predictable inflammatory outcomes.

University Of South Australia

Location: Australia

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Linkage Projects - Grant ID: LP0561810

Start Date: 12-2005

End Date: 06-2008

Amount: $165,000.00

Funder: Australian Research Council

Linkage Projects - Grant ID: LP120100606

Start Date: 01-2013

End Date: 02-2016

Amount: $183,000.00

Funder: Australian Research Council

Discovery Projects - Grant ID: DP180100520

Start Date: 07-2018

End Date: 12-2021

Amount: $303,931.00

Funder: Australian Research Council

Discovery Projects - Grant ID: DP220103543

Start Date: 07-2022

End Date: 06-2025

Amount: $529,846.00

Funder: Australian Research Council

Special Research Initiatives - Grant ID: SR180200051

Start Date: 04-2020

End Date: 04-2020

Amount: $497,638.00

Funder: Australian Research Council

Discovery Projects - Grant ID: DP150104212

Start Date: 01-2015

End Date: 12-2020

Amount: $329,900.00

Funder: Australian Research Council

Linkage Infrastructure, Equipment And Facilities - Grant ID: LE170100241

Start Date: 07-2017

End Date: 06-2018

Amount: $480,000.00

Funder: Australian Research Council

Linkage Projects - Grant ID: LP160100633

Start Date: 08-2016

End Date: 08-2019

Amount: $362,000.00

Funder: Australian Research Council