华支睾吸虫成虫抗原和排泄分泌产物对T细胞的作用研究  

Effects of crude antigen and excretory-secretory products of Clonorchis sinensis on T cells

作　　者：张波[1] 张蓓蓓[1] 程晓丹[1] 华慧[1] 于倩[1] 颜超[1] 汤仁仙[1] 郑葵阳[1] ZHANG Bo ZHANG Bei-bei CHENG Xiao-dan HUA Hui YU Qian YAN Chao TANG Reng-xian ZHENG Kui-yang(Jiangsu Key Laboratory of Immunity and Metabolism, Department of Pathogenic Biology and Immunology, Xuzhou Medical College, Xuzhou 221004, China)

机构地区：[1]江苏省免疫与代谢重点实验室徐州医科大学病原生物学与免疫学教研室/感染与免疫实验室,徐州221004

出　　处：《中国人兽共患病学报》2017年第6期491-494,共4页Chinese Journal of Zoonoses

基　　金：国家自然科学基金(No.81171590;81572019);家畜疫病病原生物学国家重点实验室开放课题(No.SKLVEB2013KFKT005);江苏高校优势学科建设工程资助项目(PAPD)联合资助~~

摘　　要：目的观察华支睾吸虫成虫抗原(Crude antigen,CA)、排泄/分泌产物(excretory-secretory products,ESPs)对T细胞的作用。方法体外分离小鼠骨髓细胞诱导分化为未成熟骨髓树突状细胞(immature DC,iDC);磁珠分选仪分选小鼠脾脏细胞的初始CD4+T细胞;流式细胞术检测DC、CD4+T细胞的纯度;抗原刺激DC细胞,实验分为PBS阴性对照组,LPS阳性对照组,CA和ESPs刺激组;负载抗原后的DC细胞与分选的CD4+T细胞共培养72h;Real time-PCR检测T-bet、GATA3mRNA的相对表达量;ELISA检测细胞培养上清中IFN-γ、IL-4细胞因子的表达量。结果与PBS组相比,ESPs刺激组Tbet、GATA3mRNA表达水平升高(P<0.05),而CA刺激组T-bet、GATA3 mRNA表达水平差异不显著;与PBS组相比,ESPs刺激组细胞因子IFN-γ、IL-4的含量均升高(P<0.05),CA刺激组仅IFN-γ的分泌增高(P<0.05),IL-4无明显变化。结论 CA可能诱导宿主产生Th1型免疫应答,ESPs可能诱导宿主产生Th1型、Th2型免疫应答。We investigated the roles of the crude antigen（CA）of Clonorchis sinensis and excretory-secretory products（ESPs）in the polarization of Th1 and Th2 cells.Bone marrow-derived cells were generated from BALB/c mice and isolated into immature DCs;immature DCs were then treated with either CA（CA stimulated group）,ESPs（ESPs stimulated group）,LPS（positive control group）or PBS（negative control group）for 24 hours.Then the CD4-＋T cells were isolated from mouse spleen by using anti-mouse-CD4 Microbeads,and further cocultured with stimulated DCs for another 72 hours.The purities of DCs and CD4-＋T cells were evaluated by flow cytometry and the expressing levels of T-bet mRNA and GATA-3mRNA were detected by real-time PCR.ELISA was used to detect the levels of IFN-γand IL-4 cytokines in the supernatant.mRNA levels of Tbet and GATA-3in the ESPs group were higher than those in PBS-stimulated group（P〈0.05）.The concentrations of IFN-γand IL-4 cytokines in the culture were increased in the ESPs group,compared with PBS stimulated group（P〈0.05）.IFN-γbut not IL-4 was increased in CA group（P〈0.05）.The results implied that CA might play a role in Th1 type immune response,and ESPs likely play roles in both Th1 and Th2 immune responses.

关 键 词：华支睾吸虫 DC细胞 CD4+细胞 成虫抗原 排泄/分泌产物 

分 类 号：R383[医药卫生—医学寄生虫学]