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目的探讨含有未甲基化“胞嘧啶-磷酸二酯键-鸟嘌呤(CpG)基序”的寡脱氧核糖核苷酸(CpG ODN)致敏人外周血来源树突状细胞(DC)在卵巢癌免疫治疗中的作用。方法应用 CpGODN2006联合肿瘤相关抗原 CA125体外冲击致敏 DCs,流式细胞技术检测 DC 膜表面 CD_(1α)、CD_(83)、CD_(86)和人白细胞 DR 抗原(HLA-DR)的表达,用 ELISA 测定 DC 培养上清液中白细胞介素(IL)-12的分泌水平;四唑盐(MTT)比色试验法检测活化 DCs 对细胞毒性 T 淋巴细胞(CTL)杀伤 OVCAR-3卵巢癌细胞株作用的影响。结果 CpG ODN2006联合 CA125在体外可明显激活人外周血来源 DCs,DC表面分子 CD_(83)、CD_(86)和 HLA-DR 表达的阳性细胞百分率[(85±4)%、(87±12)%和(92士7)%]明显高于未致敏组[(19±4)%、(67±9)%和(63±6)%](P<0.01);联合致敏后 DCs 培养上清液中白细胞介素(IL)-12分泌水平为(467±84)ng/L,与未致敏组[(60±9)ng/L]和单纯 CA125致敏组[(97±16)ng/L]相比差异有统计学意义(P<0.01);联合刺激后的 DCs 可以促进 T 淋巴细胞的增殖,增殖活性显著高于未致敏组和单纯 CA125致敏组(P<0.01),且相同效靶比下所诱导的 CTLs 对OVCAR-3卵巢癌细胞的杀伤活性显著强于未致敏组、单纯 CA125致敏组和单纯 CpG ODN 致敏组(P<0.01)。结论 CpG ODN 联合 CA125体外致敏 DC 可诱导产生对 OVCAR-3卵巢癌细胞的免疫杀伤作用,是一种临床应用前景良好的肿瘤免疫治疗方法。
Objective To investigate the effect of dendritic cells (DCs) primed by oligodeoxyribonucleotides (CpG ODNs) containing unmethylated CpG motifs, The role of immunotherapy in ovarian cancer. Methods CpG ODN2006 combined with tumor antigen CA125 was used to sensitize DCs in vitro. The expression of CD_ (1α), CD_ (83), CD_ (86) and human leukocyte antigen (HLA) DR on DCs was detected by flow cytometry. ELISA was used to measure the secretion of interleukin (IL) -12 in DC culture supernatant. MTT colorimetric assay was used to detect the cytotoxic T lymphocytes (CTLs) killing OVCAR-3 ovarian cancer cell lines Effect of the effect. Results CpG ODN2006 combined with CA125 could obviously activate the expression of CD 83, CD 86 and HLA-DR positive cells in human peripheral blood DCs in vitro [(85 ± 4)%, (87 ± 12) (19 ± 4)%, (67 ± 9)%, and (63 ± 6)%] (P <0.01). The combination of DCs The level of interleukin (IL) -12 secretion in the supernatant was (467 ± 84) ng / L, which was significantly higher than that in the non-sensitized group [(60 ± 9) ng / L) ) ng / L compared with the control group (P <0.01). The DCs stimulated by the combined stimulation could promote the proliferation of T lymphocytes, and the proliferative activity was significantly higher than that of the non-sensitized and CA125 sensitized groups (P <0.01) ), And the cytotoxic activity of CTLs induced by the same target ratio was significantly higher than that of the non-sensitized group, CA125-sensitized group and CpG ODN-sensitized group (P <0.01). Conclusion CpG ODN combined with CA125 sensitized DC in vitro can induce the immunosuppressive effect on OVCAR-3 ovarian cancer cells. It is a promising tumor immunotherapy for clinical application.