趋化因子RANTES快速募集DCs及DCs疫苗的抗结肠癌作用

目的 观察RANTES促进小鼠外周血Des前体细胞动员及DCs疫苗对结肠癌细胞的体外杀伤作用.方法 C57BL/6J(B6)小鼠静脉注射RANTES,不同时间间隔(0、4、8、16、24、48、72 h)采集外周血分离单个核细胞(PBMNCs),通过流式细胞仪分选出F4/80-B220-CD11c+细胞并其进行检测.反复冻融法制备结肠癌可溶性抗原,将其与RANTES动员的DCs共同培养,制备成DCs疫苗以激活T细胞,MTT法检测活化的T细胞在体外对结肠癌细胞的杀伤作用.γ干扰素(IFNγ)酶联免疫吸附试验(ELISA)试剂盒检测IFNγ的分泌情况.结果 B6小鼠外周血中F4/80-B220-CD11c+细胞数量随着注射时间的延长逐渐增多,大约在24 h达到高峰,占PBMNCs(13.45±1.25)%.新鲜分离的F4/80-B220-CD11c+细胞为DCs前体细胞.负载结肠癌抗原的DCs激活的T细胞表现出对结肠癌细胞的特异性杀伤作用,产生高水平的IFNγ(1595.00±38.03)ng/L,而对B16黑色素瘤细胞没有杀伤作用,不产生高水平的IFNγ(175.44±6.55)ng/L.结论 RANTES动员的DCs在体外可以诱导出针对结肠癌细胞的特异性杀伤作用.

Mobilization of dendritic cell precursors into circulation by administration of RANTES and Debased anti- colon cancer cells efficacy

Objective To investigate mobilization of dendritic cell precusor cells into the circula-tion by adiministration of RANTES, furthermore and DC-based anti- colon cancer cells efficacy ex vivo. Methods C57BL/6J (B6) mice were injected with RANTES. Peripheral blood mononuclear cells (PBMNCs) were prepared from peripheral blood at different time points (0,4,8,16,24,48 and 72 h).F4/80-B220-CD11c + cells were sorted from these PBMNCs by FACS and evaluated. DCs mobilized by RANTES injection were loaded with colon cancer antigen obtained by frozen and thawed method. The stim-Mated DCs vaccination induced T lymphocytes, and the killing effect of T cells to colon cancer ceils was assayed by MTT. IFNγ production was determined with the IFNγ ELISA kit. Results The number of F4/80-B220-CD11c + cells in PBMNCs of B6 mice was increased with time and reached the peak at approxi-mately 24 h [(13.45±1.250% of PBMNCs)] after RANTES injection. Freshly isolated F4/80-B220-CD11c + cells were DCs precusor cells. T lymphocytes stimulated with DCs loaded colon cancer antigen showed the specific killing effect on colon cancer cells and produced high levels of IFNγ (1595±38.03ng/L). However,they did not showed specific killing effect on BI6 melanoma cells and the levels of IFNγwas low (175.44±6.55 ng/L). Conclusion DCs mobilized by RANTES injection can induce specific CTL to colon cancer cells.