HLA-A~*2402 is one of the most frequent HLA-A allele in Asian population.To construct HLA-A~*2402-peptide tetramers,the transmembrane and intracellular segments of HLA-A~*2402 cDNA were replaced with BSP sequence to form a fusion gene of sHLA-A~*2402-BSP.The sHLA-A~*2402-BSP fusion protein and β2m were high-level expressed as insoluble aggregates in E.coli,and refoided to form an HLA-A~*2402-peptide monomeric complex by dilution method in the presence of an antigenic peptide.The HLA-A~*2402-peptide monomeric complex was biotinated and tetramized to prepare HLA-A~*2402-peptide tetramer.Then using the HLA-A~*2402-peptide tetramers to detect antigen-specific cytotoxic T lymphocyte(CTL)induced by artificial antigen presenting cell (aAPC)in vitro.The results showed that HLA-A~*2402-peptide tetramer was prepared correctly,and functional in detecting antigen-specific CTL in vitro,HLA-A~*2402-peptide monomeric and its multimeric complexes are expected to provide a powerful tool for studying mechanisms of immune-related diseases in Asian populations. Cellular & Molecular Immunology.2005;2(2):145-149.
IDDM results from pancreatic beta cell destruction by islet-reactive T cells,a process that involves beta cell apoptosis.Fas-FasL pathway plays a major role in pancreatic β cell death.Fas-associated death domain protein (FADD),the component of the tumor necrosis factor receptor type 1(TNFR1) and Fas signaling complexes,is involved in TNFR1-and Fas-induced apoptosis.Inhibiting the function of FADD will lead to blocking downstream apoptosis signal,which protects pancreatic β cells from destruction by Fas-FasL pathway.In this study we constructed eukaryotic expressing vector of fusional protein FADDdel-GFP named pFADDdel-GFP. After pFADDdel-GFP was transfected into NIT,the expression of FADDdel-GFP in NIT was detected by fluorescence microscopy and the resistance of NIT transfected with pFADDdel-GFP to cytotoxicity mediated by special T cells was detected by FACS and MTT.The results showed that NIT modified by pFADDdel-GFP obviously resisted cytotoxicity mediated by special T cells.Therefore,it may be useful in the prevention or treatment of IDDM by intervening Fas-FasL pathway.Cellular & Molecular Immunology.2004;1(5):383-386.
The main approach to reduce graft rejection has been focused on the development of immunosuppressive agents at present.Although these strategies have reportedly reduced graft rejection,there has been a reciprocal increase in more severe immunosuppression and lethal infections,as well as severe side effects.Blockade of costimulatory T cell response has been proved as one of useful strategies to reduce graft rejection.Furthermore, it has been shown that infusion of dendritic cells(DCs)with a potent negative regulatory ability for T cells could prolong allograft survival.In this study mouse DCs(mDCs)were transfected with the recombinant plasmid pcDNA3.0 containing mouse inducible costimulator-Ig(mICOS-Ig) cDNA by electroporation.The transient expression of mICOS-Ig in mDC could be detected by ELISA and SDS-PAGE.Mouse ICOS-Ig fusion protein expressed in mDC and mICOS-Ig gene-modified mDC could inhibit lymphocyte proliferation in mixed lymphocyte culture(MLC)in vitro.Furthermore,mICOS-Ig gene-modified mDC could inhibit lymphocyte proliferation in recipient mice.These results suggested that mICOS-Ig gene-modified mDC exerted inhibitory effects on T cells,and might be suitable for treatment or prevention of graft rejection and immunopathologic diseases.Cellular & Molecular Immunology.2004;1(2):153-157.
