树突状细胞负载肝癌抗原肽疫苗体外诱导特异性免疫学反应的研究

目的：研究树突状细胞(dendritic cell，DC)负载肝癌抗原肽EPVTKAEML体外诱导特异性CTL的能力及其抑癌效果。方法：用顺序特异引物聚合酶链反应技术(PCR—SSP)选择HLA—B7表型供者，从脾组织中分离、培养DC-EPVTKAEML特异性CTL。用^51Cr释放法检测CTL的杀伤活性，并用抗HLA-1分子单抗(mAb)进行杀伤抑制实验。结果：找到4例HLA-B7杂合子供者，用DC负载HLA-B7限制的抗原肽EPVTKAEML可诱导特异性CTL反应，对肝癌细胞HHCC有较强的杀伤作用。结论：DC负载抗原肽EPVTKAEML在体外可诱发较强的特异性免疫反应。

Study on immunoreaction induced by DCs loaded with hepatocarcinoma antigen peptide in vitro

AIM: To investigate the potency and antitumor effect of specific cytotoxic T lymphocytes induced by hepatocarcinoma antigen peptide EPVTKAEML-loaded dendritic cells (DCs). METHODS: The healthy donors of HLA-B7 genotype were selected by PCR-SSP. Isolated and cultured DCs from HLA-B7 donors' spleen tissues were loaded with the peptide EPVTKAEML from hepatocarcinoma cell line HHCC. The specific CTL reaction and inhibitory effect of anti-HLA-I molecule mAb on CTL cytotoxicity were detected in-vitro by (51)Cr-release assay. RESULTS: 4 donors with HLA-B7 genotype were found. Their spleen T lymphocytes could be induced to proliferate and differentiate into specific CTLs by DCs loaded with the peptide. Antibody-blocking test demonstrated that the cytotoxity of CTLs was strictly limited by MHC-I molecule. CONCLUSION: EPVTKAEML-loaded DCs can induce efficient and specific antitumor immune reaction in-vitro.