【摘要】 目的检测来源于HCA587的HLA-A2限制性表位肽免疫HLA-A2转基因小鼠后产生细胞应答的水平,筛选出体内具有免疫原性的HLA-A2限制性表位肽。方法将来源于HCA587的候选HLA-A2限制性肽与MHC-Ⅱ类限制性肽HBV-core128-140、不完全弗氏佐剂、Cp G ODN1826联合应用皮下免疫HLA-A2转基因小鼠,用酶联免疫斑点实验(ELISpot)检测小鼠脾细胞表位肽特异性细胞免疫应答的水平,流式细胞术检测表位肽特异性的CD8+T细胞比例。结果在检测的4个表位肽中,p248-256诱导HLA-A2转基因小鼠产生的细胞免疫应答最强,HLA-A2-H-2Kb-p248-256四聚体染色进一步表明p248-256免疫后可产生p248-256特异性的CD8+T细胞。结论来源于HCA587的HLA-A2限制性表位肽中,p248-256具有较强的体内免疫原性。更多还原

【Abstract】 Peptide vaccines have been conceived as promising therapies for tumor-inflicted patients due totheir easy production and capability of inducing specific immune response required for defending the tumor. Duringprevious research, 4 HLA-A2-restrictd peptides had been identified as immunogenic in vivo. In this study, weaimed to testify the in vivo immunogenicity of the 4 peptides. HLA-A2 transgenic mice were vaccinated withHLA-A2 restricted peptides(peptide pool or single peptide) and MHC-Ⅱ-restricted peptide HBVcore128-140 emulsified in incomplete Freud adjuvant(IFA) subcutaneously in combination with repeated subcutaneous injectionof Cp G ODN1826 at the adjacent location. 10 days after the third peptide vaccination, the peptide-specific immuneresponse was evaluated by ELISpot assay, and the presence of peptide-specific CD8+T cells capable of recognizingthe MHC-peptide tetramer was detected by flow cytometry assay. Our results showed that among the 4 candidateHLA-A2 restricted peptides, the immune response elicited by p248-256 was superior to that of the other 3 peptides, andHLA-A2-H-2Kb-p248-256 tetramer assay indicated the presence of p248-256-specific CD8+T cells in p248-256-vaccinatedmice. These results suggest that p248-256 is the most immunogenic peptide in vivo, and can be explored as potentialtumor peptide vaccine in future clinical study.更多还原