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目的研究华支睾吸虫成虫虫体抗原(CA)致敏树突状细胞(DCs)诱导免疫应答的类型。方法利用rm-GMCSF诱导分化BALB/c小鼠骨髓细胞获得DCs,倒置显微镜和透射电镜观察DCs形态;CA致敏DCs,检测IL-12p70水平;将致敏后的DCs和淋巴细胞共培养,用CCK-8试剂盒检测淋巴细胞增殖,用ELISA试剂盒检测IFN-γ和IL-4水平。结果倒置显微镜和透射电镜下见典型的DCs,CA致敏的DCs分泌高水平IL-12p70;共培养后,淋巴细胞明显增殖,分泌的IFN-γ水平显著升高,IL-4水平与其他各组比较差异无统计学意义(P>0.05)。结论CA刺激后DCs被活化,诱导的免疫应答向Th1方向极化,该应答与华支睾吸虫感染早期的保护性免疫密切相关。
Objective To study the types of immune responses induced by dendritic cells (DCs) of adult cells of Clonorchis sinensis. Methods DCs were isolated from BALB / c mouse bone marrow cells by rm-GMCSF. Morphology of DCs was observed under inverted microscope and transmission electron microscope. DCs were detected by CA-primed DCs to detect the level of IL-12p70. DCs and lymphocytes were co-cultured with Lymphocyte proliferation was detected by CCK-8 kit and IFN-γ and IL-4 levels by ELISA kit. RESULTS: Typical DCs were seen under inverted microscope and transmission electron microscope. CA-sensitized DCs secreted high levels of IL-12p70. After co-culture, lymphocytes proliferated significantly and secreted IFN-γ levels were significantly increased. There was no significant difference between the two groups (P> 0.05). Conclusions After stimulated by CA, DCs are activated and the induced immune responses are polarized toward Th1. This response is closely related to the early protective immunity of Clonorchis sinensis infection.