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

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.