目的: 探讨紫杉醇联合顺铂诱导凋亡的卵巢癌细胞被DCs交叉提呈后能否促进免疫应答。方法：常规贴壁法刺激正常人外周血单核细胞分化诱导为DCs，6 d后将DCs和紫杉醇联合顺铂体外诱导的凋亡人卵巢癌细胞HO8910共同培养（凋亡组），并以冻融细胞共培养DCs组(冻融组)或单独DCs组(空白组)作对照，以激光共聚焦扫描和流式细胞仪检测不同培养组DCs的分化和成熟程度。分别将各组成熟DCs与同一来源的磁珠分选的CD8+T细胞共培养, 3HTdR掺入法检测其增殖能力；LDH释放反应检测CTL对肿瘤细胞的杀伤能力；同时应用ELISPOT图像分析仪检测致敏后CD8+T细胞INFγ的分泌能力。结果：(1)紫杉醇联合顺铂诱导的凋亡卵巢癌细胞可被DCs有效吞噬，并促进DCs的成熟及其抗原提呈作用；(2) 3HTdR检测显示凋亡肿瘤细胞能诱导CD8+T细胞增殖（P<0.05）；(3)LDH释放实验与ELISPOT图像分析均证明凋亡肿瘤细胞诱导的CTL杀伤活性显著强于冻融组和空白组细胞（P<0.01）。结论：紫杉醇联合顺铂诱导的凋亡卵巢癌细胞具有较强的免疫原性，可促进DCs分化和成熟，进一步促进CD8+T细胞增殖、分泌与杀伤功能。

Objective: To explore whether apoptotic ovarian cancer cells induced by chemotherapy drugs paclitaxel and cisplatin can be crosspresented by dendritic cells (DCs) and enhance immune responses. Methods: DCs were induced from peripheral blood monocytes cells by GMCSF/IL4 for 6 d, then they were stimulated with either apoptotic ovarian cancer HO8910 cells, frozenthawed HO8910 cells or control cells for 4 h. Their surface markers and phagocytotic ability were detected by flow cytometry and confocal microscopic scanning assay, respectively. DCs of different groups were cultured with CD8+ T cells isolated by magnetic cell sorting, and the ability of DCs to activate CD8+ T cells was evaluated by 3HTdR, the activity of CTL to kill tumor cells was evaluated by LDH. Production of IFNγ by CD8+ T cells was measured by ELISPOT. Results: Apoptotic ovarian cancer cells induced by chemotherapy drugs paclitaxel and cisplatin could be phagocytized by DCs, which subsequently promoted the maturation and antigen presenting ability of DCs. Apoptotic ovarian cancer cells implused DCs significantly promoted proliferation of CD8+ T cells compared with that of control cells (P＜0.05）; they also significantly increased the cytotocity of CTL to kill tumor cells compared with that of frozenthawed HO8910 cells and control cells as detected by LDH and ELISPOT（all P＜0.01). Conclusion: Apoptotic ovarian cancer cells induced by paclitaxel and cisplatin exhibit strong immunogenicity and enhanced ability to promote DCs maturation and antigen presentation, subsequently enhance CD8+ T cell proliferation and promote their ability to secrete IFNγ and kill tumor cells.

上海市教育委员会科研项目（No.061012）