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目的探讨构建K-ras(12-Val)树突状细胞(DC)疫苗体外诱导抗胰腺癌特异性免疫反应的可行性。方法采用健康人外周人单核细胞(PBMC)体外刺激分化的DC,负载K-ras(12-Val)多肽后诱导成熟,流式细胞仪测定其细胞表型;与PBMC混合培养扩增特异性细胞毒性T淋巴细胞(CTL),MTT法测定CTL对胰腺癌细胞和正常细胞的免疫杀伤活性。结果负载K-ras(12-Val)多肽的DC成熟后能较好表达突变位点并能有效地刺激T淋巴细胞活化,其诱导产生的CTL对表达该突变的胰腺癌细胞有较好的免疫杀伤作用,对正常组织无损伤作用。结论利用K-ras(12-Val)DC疫苗能成功诱导出较强的抗胰腺癌免疫反应,且具有高度特异性。
Objective To investigate the feasibility of inducing anti-pancreatic cancer-specific immune response in vitro by constructing a K-ras (12-Val) dendritic cell (DC) vaccine. Methods DCs stimulated by peripheral blood mononuclear cells (PBMCs) were cultured and stimulated with 12-Val in vitro. The cell phenotypes were determined by flow cytometry. Cytotoxic T lymphocytes (CTLs) and CTLs were used to determine the immunosuppressive activity of CTLs against pancreatic cancer cells and normal cells. Results The DCs loaded with K-ras (12-Val) could well express the mutated sites and stimulate the activation of T lymphocytes effectively. The CTLs induced by the recombinant CTLs had good immunity to the pancreatic cancer cells expressing the mutation Killing effect, no damage to normal tissue. Conclusion K-ras (12-Val) DC vaccine can induce a strong anti-pancreatic cancer immune response with high specificity.