ORCID Profile
0000-0003-0323-637X
Current Organisation
Monash University
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Publisher: Elsevier BV
Date: 07-2020
Publisher: Elsevier BV
Date: 06-2023
Publisher: Elsevier BV
Date: 2019
DOI: 10.1016/J.MICPATH.2018.11.029
Abstract: It is well established that the current problem of tuberculosis (TB) can be combated by overcoming the drawbacks of the currently available BCG vaccine. This would involve incorporation of antigens that can control TB at all stages including the dormant phase which is generally ignored. Hence, DosR regulon proteins, which are expressed in latent infection, could prove to be very good vaccine candidates as they can possibly target the silent but most predominant form of TB infection. In the present study, the immune response to two DosR proteins Rv2627 and Rv2628 has been studied in PBMCs derived from normal in iduals, TB patients and healthy contacts of TB patients. It was found that these antigens were capable of stimulating a strong IFN-γ+ T cell response along with accentuation of memory T cells and other protective cytokines such as IL-2 and IL-17. At the same time these proteins decreased the frequencies of immune-suppressor regulatory T cells in in vitro stimulation of PBMC from both patients and their contacts. Considering all these facts together, we suggest Rv2627 and Rv2628 to be one of the extremely promising candidates for incorporation into a post exposure subunit vaccine against TB.
Publisher: Medknow
Date: 03-2016
DOI: 10.1016/J.IJMYCO.2015.10.005
Abstract: There is an urgent need for a more effective vaccine against Mycobacterium tuberculosis (Mtb). Although CD4+ T cells play a central role in host immunity to Mtb, recent evidence suggests a critical role of CD8+ T cells in combating Mtb. In the present study, we have predicted HLA antigen class I binding peptides of DosR operon using an in-silico approach. This method is useful as an initial computational filtration of probable epitopes based on their binding ability and antigenicity. CD8+ epitopes were predicted by software NetMHC 3.4 and BIMAS. Self-peptides were found and excluded by indigenously developed Perl script. Antigenicity of promiscuous peptides was predicted using a VaxiJen server. The top VaxiJen scoring antigenic peptides were docked to globally relevant HLA allele using CABS dock and Hex program. A total of 1436 overlapping nonamer peptides were generated which gave 46 promiscuous epitopes, 25 were predicted to be antigenic. Rv2627 epitope "SAFRPPLV" which gave the highest Vaxijen score of 1.9157 and showed binding to all the three HLA loci. The top VaxiJen scoring antigenic peptides were docked and had significant interactions with residues of the HLA class I molecule indicating them to be good cytotoxic T lymphocyte epitopes. Our study has generated several promiscuous antigenic peptides capable of binding to major histocompatibility complex class I with high affinity. These epitopes can become part of a postexposure multivalent subunit vaccine upon experimental validation.
Publisher: Elsevier BV
Date: 2021
Publisher: Elsevier BV
Date: 2023
Publisher: Wiley
Date: 03-2021
DOI: 10.1002/CPZ1.92
Abstract: This article describes the purification of HLA‐bound peptides and their subsequent sequencing by mass spectrometry. These methods can be used for both HLA class I and class II molecules and can be adapted to different species depending on the availability of specific antibodies. Peptides can be successfully isolated from a variety of s le types, including in vitro cultured cells and primary tissues. The method involves the affinity capture of HLA‐peptide complexes and separation of peptides from HLA heavy chains, followed by tailored interrogation by mass spectrometry to take into account the non‐tryptic nature of endogenously derived HLA‐bound peptides. © 2021 Wiley Periodicals LLC. Basic Protocol 1 : Preparation of immunoaffinity column Alternate Protocol 1 : Preparation of microscale immunoaffinity column Basic Protocol 2 : Generation of cell lysate and HLA immunoaffinity purification Alternate Protocol 2 : Microscale immunoaffinity purification Basic Protocol 3 : Separation of HLA peptides by reverse‐phase high‐performance liquid chromatography (RP‐HPLC) Alternate Protocol 3 : Isolation of HLA peptides using molecular weight cutoff (MWCO) filter Basic Protocol 4 : Mass spectrometry and data analysis
Publisher: Wiley
Date: 23-09-2016
DOI: 10.1111/CBDD.12840
Abstract: Tuberculosis is a global health problem especially with the emergence of drug-resistant Mycobacterium tuberculosis strains, creating an urgent need to identify new drug targets. The mycobacterial cell wall is an attractive target for chemotherapeutic agents. Gene products of mymA operon are known to be required for the maintenance of cell wall and play an important role in persistence, thus making them important drug targets. This study was undertaken to biochemically characterize the MymA as a flavin-containing monooxygenase (FMO). Our results established its enzymatic activity in vitro and found that the mycobacterial FMO requires NADPH and FAD as cofactors, similar to other characterized bacterial FMOs. The enzyme follows Michaelis-Menten kinetics to catalyze substrates such as trimethylamine and thiourea. We also propose that MymA could be one of the targets of the antituberculosis drug, isoniazid (INH), which is a cell wall inhibitor. Molecular docking studies revealed that INH targeted NADPH-binding site of the MymA. Further, experimental validation revealed that INH inhibits MymA with the IC
Publisher: Elsevier BV
Date: 09-2017
DOI: 10.1016/J.TUBE.2017.05.005
Abstract: Cell wall of Mycobacterium tuberculosis (M.tb) is a major source of immunogenic proteins that can be tested as vaccine candidates. MymA (Rv3083), a 55 kDa M.tb flavin containing monooxygenase, is involved in modification of mycolic acids during acidic shock following M.tb internalization in macrophage. In this study, we have investigated the role of this cell wall associated protein in activation of macrophages by toll like receptor (TLRs) engagement and subsequent signaling. Our results showed that MymA stimulation of THP1 cells and human monocyte derived macrophages (MDM) lead to upregulation of TLR2 and co-stimulatory molecules CD40, CD80, CD86 and HLA-DR. This upregulation is partially reduced by TLR2 blocking antibodies. The activation of macrophage following MymA stimulation also resulted in release of proinflammatory cytokines, TNF-α and IL-12. Moreover, MymA also polarized the immune response towards T
No related grants have been discovered for Boliang Wang.