CD40配基化调节小鼠树突状细胞上B7-H3分子表达的研究  

Regulatory effects of CD40 ligation on B7-H3 expression in murine dendritic cells

作　　者：陈成[1] 朱一蓓[1] 周桓[1] 瞿秋霞[1] 王勤[1] 马倩茹[1] 胡玉敏[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,215007

出　　处：《中华微生物学和免疫学杂志》2006年第3期228-231,共4页Chinese Journal of Microbiology and Immunology

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

摘　　要：目的探讨CD40配基化对小鼠骨髓来源树突状细胞上B7H3分子表达的调节作用及其生物学意义。方法采用GMCSF和IL4联合方案体外诱导小鼠髓系DC,并利用mCD40LCHO和TNFα分别刺激凋亡肿瘤细胞负载的DC制备成熟DC;采用间接免疫荧光标记法检测成熟DC上B7H3分子的表达;RTPCR检测B7H3mRNA转录水平;混合淋巴细胞反应(MLR)和B7H3单抗阻断实验分析CD40配基化的DC表面B7H3分子在T细胞活化中的作用;3HTdR掺入试验检测DC对T淋巴细胞的促增殖效应;ELISA测定各组MLR反应和DC培养上清中IFNγ的分泌水平。结果B7H3分子在DC不同分化发育阶段均有表达,CD40配基化能显著上调凋亡肿瘤细胞负载的DC中B7H3表达,TNFα激发的DC弱表达(P<0.05);阻断CD40配基化的DC上B7H3分子能抑制T细胞增殖和IFNγ的分泌;CD40配基化促进凋亡肿瘤细胞负载的DC分泌IFNγ的量也明显高于TNFα组(P<0.05)。结论体外CD40配基化DC的B7H3分子上调性表达有助于其刺激T细胞增殖和IFNγ的产生。Objective To study the regulatory effects of CD40 ligation on B7-H3 expression in murine dendritic cells and its biological significance. Methods Murine myeloid dendritic cells（DC） were generated from bone marrow in vitro using GM-CSF and IL-4, apoptotic tumor cells pulsed DC were induced further maturation by CD40L-CHO cells or TNF-α for 48 h, respectively; expressions of B7-H3 in DC was detected by FCM and mRNA of B7-H3 was detected by RT-PCR. ^3H-TdR incorporation test was used to detect the T cell proliferation stimulated by DC with or without blocking B7-H3 by McAb respectively. The concentration of IFN-γ in supematants of MLR from distinct groups was analyzed by ELISA. The concentration of IFN-γ in supernatant of DC was determined by ELISA. Results The B7-H3 was up-regulated in DC simulated with CD40L-CHO for 48 h. Differential expression of BT-H3 was impressive on DC stimulated by mCD40L-CHO cells or TNF-α. Blockade of BT-H3 on CD40 Kgation DC inhibits T cell proliferation and decreases IFN-γ secretion. CD40 ligation was a potent signal to enhance apoptotic tumor cells pulsed DC to secrete IFN-γ. Conclusion High expression of B7-H3 contributes to CD40 ligation DC to induce generation of TH 1 effectors.

关 键 词：树突状细胞 CD40 B7-H3 

分 类 号：R392[医药卫生—免疫学]