肝癌抗原肽SLIVHLNEV免疫原性研究

Immunogenicity of the Hepatocellar Carcinoma Antigenic Peptide SLIVHLNEV

肿瘤细胞抗原肽SLIVHLNEV可以与MHC-I类分子结合,由T细胞表位识别,激活细胞毒性T细胞(CTL),为肝癌免疫治疗开辟了新领域。文章通过软件DNAstar-protean与Syfpeithi分析,确定抗原肽的抗原表位。然后抗原肽刺激树突细胞,通过检测CD80,CD86的表达,观察抗原肽诱导树突细胞成熟的情况。将负载抗原肽的树突细胞与T细胞的混合淋巴细胞反应(MLR),MTT检测T细胞增殖的情况,确定最佳抗原肽浓度与最佳刺效比。以最佳抗原肽浓度与最佳刺效比刺激T淋巴细胞分化,测定T细胞表面CD8、CD4表达情况。结果表明抗原肽SLIVHLNEV不含B淋巴细胞抗原表位,而为HLA_A*02∶01型T淋巴细胞表位抗原,并可以刺激H2-Db表位,经树突细胞(APC)递呈,刺激T细胞分化为CTL,发挥细胞免疫作用。

The tumor antigenic peptide SLIVHLNEV which can be bound to MHC class I,recongnized by T lymphocytes and futher to activate the cytotoxic T cells（CTL）,opened up a new field in tumor immunotherapy.In this paper,the epitope of antigenic peptide was analyzed by the software DNAstar-protean and SYFPEITHI.Then the dendritic cells were stimulated by the antigen peptide,and the expression of CD80 and CD86 was detected to investigate the antigenic peptide-induced maturation of dendritic cell.T cells were co-cultured with dendritic cells stimulated by antigenic peptides in the mixed lymphocyte reaction（MLR）,and the proliferation of T cells was analyzed by MTT to determine the optimal concentration of the peptide and the best ratio of T cells to dendritic cells.Then,at the optimal concentration and ratio obtained above,the differentiation of T lymphocyte was stimulated and the expression level of CD8 and CD4 on the T cell surface was determined.The results showed that the antigen peptide SLIVHLNEV contains HLA_A ＊ 02∶ 01 T lymphocyte antigen epitope while none of the B lymphocyte antigen epitopes is included,and it can stimulate the H2-DB epitope,too.The expression level of CD80 and CD86 of the dendritic cells gets rised after stimulation by the antigen peptide.It proves that the tumor antigenic peptide SLIVHLNEV can stimulate the DCs＇ maturation.In MLR experiment,the antigen peptide induced the T cells proliferation after the presentation by dendritic cells（APC）.The optimal concentration of peptide is 50 μmol / L and the best ratio of T cells to dendritic cells is 51.Under this condition,T cells＇ differentiation is mainly into CTL,which is the role of cell-mediated immunity.