人卵巢癌细胞光动力学细胞裂解物负载树突状细胞对体外免疫应答的影响

Effect of immune response of dendritic cells after loaded with photodynamic therapy-generated human ovarian tumor cell lysates in vitro

目的:研究人卵巢癌细胞光动力学细胞裂解物对体外免疫应答反应的影响。方法:体外诱导培养人脐血树突状细胞(DC)。人卵巢癌细胞SKOV3分别用光动力学方法(激光照射)及冻融的方法制备细胞裂解物,与DC共培养48 h,RPMI1640与DC培养作为阴性对照,采用流式细胞仪检测DC免疫表型CD1a、CD80、CD86、HLA-DR的改变,用ELISA方法检测培养上清IL-12含量的改变。体外培养T细胞,用负载裂解物的DC诱导生成细胞毒性T细胞(CTL),MTT法检测CTL对人卵巢癌细胞的杀伤效应。结果:在体外,人卵巢癌细胞光动力学裂解物较冻融裂解物更能促进DC的成熟,表现为DC细胞表型中共刺激分子CD80、CD86上调,分泌更多的IL-12,能诱导CTL产生更强的特异性肿瘤细胞杀伤效应。结论:人卵巢癌细胞的光动力学细胞裂解物较冻融细胞裂解物在体外更能促进DC成熟,并引发CTL的特异性肿瘤杀伤效应,因此,具有更强的肿瘤疫苗样作用。

Objective ： To investigate the effect of photodynamic therapy （PDT）-generated human ovarian carcinoma cell lysates on cellular immune response in vitro. Methods：Cord blood cells as precursor were used to induce dendritic cells （DCs） differentiation . Two kinds of human ovarian carcinoma cell lysates were loaded to DCs. One was generated by PDT, the other by freeze-melt method. The co-cultured time was 48 hours. DCs cultured with RPMI1640 served as negative control. Flow cytometry was used to detect the expression of CD80, CD86, CD1a, HLA-DR. Production of IL-12 in the DCs culture supernatant was determined by enzyme linked immunoadsorbent assay （ELISA）. T cells were isolated from cord blood and co-cultured with DCs to induce cytotoxic T lymphocytes （CTLs）. Tumor killing elects of CTLs were assayed by MTT. Results： In vitro, PDT-generated tumor cell lysates induced DCs to be more mature and produced more IL-12 than freeze-melt. CTLs activated by DCs loaded with PDT-generated tumor cell lysates were more efficient in killing tumor cells. Conclusion： In vitro, PDT-generated tumor cell lysates can promote maturation of DCs and induce more efficient killing effect on tumor cells by CTLs than freeze-melt. Therefore, PDT-generated tumor cell lysates have stronger vaccine-like effect.