华支睾吸虫成虫抗原致敏树突状细胞诱导免疫应答的研究  

Dendritic cells pulsed by an antigen of Clonorchis sinensis to induce immune response

作　　者：戴其锋[1] 付琳琳[1] 刘宜升[1] 杜文平[1] 郑葵阳[1] 汤仁仙[1] 郭倩倩[1] 范宝峰[1] 张嫒嫒[1] 

机构地区：[1]徐州医学院病原生物学教研室,江苏徐州221000

出　　处：《中国病原生物学杂志》2009年第8期582-585,F0004,共5页Journal of Pathogen Biology

基　　金：江苏省高校自然科学基础研究项目(No.07KJB310122);卫生部寄生虫病预防与控制技术重点实验室和江苏省寄生虫病预防与控制高技术平台开放研究课题(No.wk008-003);江苏省血地寄防科研课题(No.X200731)

摘　　要：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 investigate how T cell polarization is altered by dendritic cells （DCs） activated with a crude antigen of Clonorchis sinensis （CA）. Methods rmGM-CSF was used to induce BALB/c mouse bone marrow cells to differentiate into DCs. The morphology of DCs was observed under an inverted microscope and transmission electron microscope （TEM）. After DCs were pulsed by CA, the supernatant was collected for IL-12p70 measurement. CA-pulsed DCs were mixed with allogeneie lymphocytes and co cultured, and the supernatant was collected to detect IFN-T and IL-4 with enzyme-linked immunosorbent assay （ELISA） ; T-cell proliferation was detected with a Cell Counting Kit-8 （CCK-8）. Results Typical morphological characteristics of DCs were apparent under the inverted microscope and TEM. CA-pulsed DCs secreted higher levels of IL-12 compared to the negative control （P〈0.05）. After the cells were co-cultured, the CA-pulsed DCs had marked T-cell proliferation （P〈0.05） and markedly higher IFN-T production compared to the negative control group （P〈0.05）, although the difference in IL-4 production was not statistically significant （P〉0.05）. Conclusion CA-pulsed DCs activate immunocompetence and induce Thl-type responses that are closely associated with C. sinensis infection in early protective immunity.

关 键 词：华支睾吸虫 成虫抗原 树突状细胞 细胞因子 

分 类 号：R383.22[医药卫生—医学寄生虫学]