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Objective To identify hepatitis C virus(HCV)-specific cytotoxicity T lymphocytes (CTL)epitopes by the combination of T epitopes prediction software and in vitro assays.Methods HCVspecific CTL epitopes were predicted by T epitope prediction software Rankpep and then candidate HCV-specific CTL epitopes were selected.Candidate HCV-specific CTL epitopes were used to stimulate PBMC of HCV-infected patients and healthy volunteers.and then enzyme-linked immunospot(ELISPOT)and intracellular cytokine staining(ICS)were used to measure the frequencies of IFN-γ-producing cells in total PBMC and the percentages of IFN-γ+CD8+T cells in total CD8+T cells,respectively.Results Five candidate CTL epitopes[NS3 450(TVPQDAVSR),NS3 594(GPTLLYRL),Ns4b 78(sMMAFSAAL),NS5a 416(SEENVSVVF)and NS5a 367(TVSSALAEL)]were used to stimulate PBMC of ten HCV-infected patients and two healthy volunteers.PBMC of seven HCV-infected patients secreted IFN-γ while PBMC of healthy volunteers did not produce IFN-γ.The frequencies of peptide-specific IFN-γ-producing cells ranged from 5 to 36 SFC/105 PBMC and the percentages of peptide-specific IFN-γ+CD8+T cells ranged from 0.02%-0.25%.Conclusion Resuhs of ELISPOT assay and ICS assay confirm that these five peptides NS3 450,NS3 594,NS4b 78,NS5a 416 and NS5a 367 are identified as Hovel HCV-specific CTL epitopes.
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Objective:To explore the immunogenecity of four Dengue virus-specific peptides in mice.Methods:Each peptide of four Dengue virus-specific peptides (C45-57KLVMAFIAFLRFL,E396-408SSIGKMFEATARG,NS323-35YRILQRGLLGRSQ,and NS3141-155NREGKIVGLYGNGVV) was used to immunize BALB/c mice and C57BL/6 mice,respectively.Three weeks later,mice were killed and splelocytes were prepared.Splelocytes were stimulated by the same peptide and intracellular cytokine staining (ICS) assay was used to detect the percentages of peptide-specific IFN-? or IL-4 producing CD4+ T cells in total CD4+ T cells.Results:Peptide C45-57 induced peptide-specific IFN-?+ CD4+ T cells(0.72%?0.04% vs 0.04%?0.02%,P