CD40配基化的肿瘤特异性树突状细胞介导Th1细胞分化的研究  

Study of CD40 ligation tumor specific dendritic cells on mediating Th1 cells polarization

作　　者：陈成[1] 朱一蓓[1] 瞿秋霞[1] 周桓[1] 王勤[1] 於葛华[1] 李文香[1] 张学光[1] 

机构地区：[1]苏州大学医学生物技术研究所江苏省医学临床免疫学重点实验室,苏州215007

出　　处：《中国免疫学杂志》2006年第2期123-127,共5页Chinese Journal of Immunology

基　　金：国家重点基础研究发展规划(973计划)资助项目(2001CB510003);国家自然科学基金重点项目(30330540)

摘　　要：目的:探讨CD40配基化的肿瘤特异性DCs在介导Th1细胞分化中的作用。方法:采用GM-CSF和IL-4联合方案体外诱导小鼠髓系DCs,并利用mCD40L-CHO和TNF-α分别刺激凋亡肿瘤细胞负载的DCs制备DCs瘤苗;3H-TdR掺入试验检测DCs对T淋巴细胞的促增殖效应;ELISA测定细胞培养上清中IL-10、IFN-γ、IL-12的含量;胞内染色和流式细胞术检测经成熟DCs活化的T细胞中CD4+IFN-γ+T和CD4+IL-4+T的比例。结果:体外刺激T细胞增殖能力在CD40配基化DCs组最高(P<0.05),CD40配基化DCs能更有效地促进活化T细胞分泌IFN-γ和介导CD4+IFN-γ+T细胞的分化(P<0.05),同时,CD40配基化DCs分泌IL-12的量也明显高于TNF-α组(P<0.05)。结论:CD40配基化的肿瘤特异性DCs体外能有效介导Th1细胞的分化。Objective：To investigate the role of CD40 ligation tumor specific dendritic cells in Thl cells polarization. Methods：Mouse myeloid DCs were generated from bone marrow in vitro using GM-CSF and IL-4, apoptotic tumor cells pulsed DCs were induced further maturation by mCD4OL-CHO cells and TNF-α for 48 h, respectlvely;^3H-thymidine incorporation test was used to detect the T cell proliferation stimulated by mature DCs. The concentrations of IL-10 and IFN-γ in supernatants of MLR from dendritic celldriven T cells activation were analyzed by ELISA. Intracellular staining and FCM were used to detect the percent of CD4^＋IFN-γ^＋T cells and CD4 ^＋ IL-4 ^＋T cells in T cells activated by mature DCs. The concentration of IL-12 in supernatant of DCs was determined by ELISA. Results：In vitro proliferation of T cells induced by CD40 ligatlon DCs were more powerful than those in other groups （P 〈 0. 05 ）. CD40 ligation DCs induced more IFN-γ-producing CD4 ^＋ T cell than TNF-α stimulated DCs. CD40 ligation DCJT cultures showed a pronounced increase in the production of IFN-γ and a modest enhancement of IL-10 secretion compared with TNF-α stimulated DC/ T cultures. Moreover, CD40 ligation was a potent signal to enhance DCs to secrete IL-12. Condusion：CD40 ligation DCs were potent to mediate Thl polarization in vitro.

关 键 词：树突状细胞 CD40 TH1细胞 

分 类 号：Q254[生物学—细胞生物学] R392.1[医药卫生—免疫学]