负载卵巢癌冻融抗原对树突状细胞分泌细胞因子的影响

目的探讨负载卵巢癌冻融抗原对树突状细胞分泌细胞因子的影响。方法采用反复冻融法获得卵巢癌细胞株SKOV3细胞抗原,联合应用重组人粒细胞-巨噬细胞集落刺激因子、重组人白细胞介素-4和重组人肿瘤坏死因子-α,体外诱导人外周血CD14^＋单核细胞为成熟DCs并负载卵巢癌肿瘤抗原;采用ELISA法检测培养细胞上清液中分泌IL-12p70、IFN-γ和IL-10含量,评价非成熟DCs和成熟DCs以及肿瘤抗原负载DCs和未负载肿瘤抗原DCs激活的CTL分泌细胞因子的能力。结果联合应用重组人的GM-CSF、IL-4和TNF-α可在体外诱导出成熟DCs;经ELISA方法检测,不同状态下的DCs分泌IL-12p70、IFN-γ的量不同,成熟DCs较非成熟DCs有更强的分泌IL-12p70、IFN-γ的能力（P〈0.05）,负载抗原DCs激活的CTL较未负载抗原DCs激活的CTL有更强的分泌IL-12p70[（182.89±4.57）pg/ml和（99.76±5.42）pg/ml,P〈0.01]和IFN-γ的能力[（102.11±5.95）pg/ml和（68.29±5.04）pg/ml,P〈0.01]。而成熟DCs分泌IL-10的能力与非成熟DCs相比,差异无统计学意义（P〉0.05）。负载抗原后DCs激活的CTL与未负载抗原DCs激活CTL上清液中IL-10的浓度相比,两者之间的差异无统计学意义（P〉0.05）。结论细胞因子的分泌量受DCs的成熟状态及某些刺激信号的影响,卵巢癌冻融抗原是其刺激信号之一。

Effect of the Ovarian Carcinoma Cells Lysate on the Cytokine-Secreting Capacity of DCs

Objective To detect the effect of the SKOV3 ovarian carcinoma cells lysate on the cytokine-secreting capacity of the dendritic cells (DCs).Methods Soluble antigens were obtained from SKOV3 ovarian carcinoma cells lysate by cycling freezing-thawing. DCs were generated from human peripheral blood monocytes,which were isolated from peripheral blood of healthy donors by ficoll-hypaque gradient centrifugation and purified by using MACS CD14 isolation kit. To induce DCs,the purified CD14+ cells were cultured in t...