人脐血来源树突状细胞体外扩增及诱导特异性抗宫颈癌免疫应答

In Vitro Generation of Dendritic Cells from Cord Blood and Induction of Specific Antitumor Immunity in Cervical Carcinoma Cells

目的:研究脐血来源树突状细胞(DC)体外扩增及诱导特异性抗宫颈癌细胞的免疫效应。方法:自人脐血分离单核细胞诱导DC,制备宫颈癌细胞系CaSki冻融物作为抗原负载DC。ELISA法检测成熟DC上清中IL-12的含量,混合淋巴细胞反应(MLR)测定DC体外刺激T细胞增殖的能力,MTT法检测抗原负载DC激活的CTL对宫颈癌细胞CaSki和HeLa的杀伤作用。结果:与未经抗原负载的DC相比,抗原负载的DC刺激同种异体T细胞增殖和IL-12分泌的能力均有提高(P<0·05),激活的CTL对CaSki细胞有特异性杀伤,而此CTL对HeLa细胞无特异性杀伤。结论:宫颈癌细胞CaSki冻融抗原负载DC激活的CTL可诱导出特异性杀伤宫颈癌细胞的免疫效应。

Objectlve;To investigate the ability of generation of dendritic cell （DC） from precursor in cord blood and inducing specific antitumor immune response. Methods： Monocytes were isolated from cord blood and cultured to differentiate into DC induced by cytokines. DCs were loaded with freeze-thaw antigen of CaSki cells. IL-12 production in DC suspension was detected using ELISA method,and DC function was evaluated their ability to induce proliferation of allogenaic T cells by mixed lymphocyte reaction （MLR） assay. The cytotoxicity to CaSki cells and Hela cells of CTL activated by dendritic cells loaded with antigen was detected by MTT method. Results：Antigen-loaded DC had strong ability of inducing proliferation of allogeneic T cells and more secretion of IL-12 than that of DC without loading antigen. And activated CTL had specific antitumor effect to CaSki cells, not to HeLa. Conclusions： CTL activated by DC loaded with freeze-thaw antigen of CaSki cells could induce most potent specific cytotoxicity to cervical carcinoma cells in vitro.