CD40培养体系的建立及B淋巴细胞的体外培养

采用自行研制成的功能性抗人ＣＤ４０单克隆抗体、转导ＣＤ３２的Ｌ细胞（ＬＣＤ３２）和细胞因子建立体外研究人Ｂ淋巴细胞增殖、生长和分化的ＣＤ４０系统。方法：①流式细胞仪分析ＣＤ４０分子的表达；②在ＣＤ４０激发型单抗（克隆５Ｃ１１）与ＬＣＤ３２共培养中，加入ＩＬ－４及其它细胞因子，组成ＣＤ４０培养系统，观察人扁桃体来源Ｂ细胞的体外生存和增殖等生物学效应。结果：①ＣＤ４０分子表达于人外周血、扁桃体来源的Ｂ细胞及人多发性骨髓瘤（ＭＭ）细胞株ＸＧ２和人Ｂ淋巴瘤细胞株Ｄａｕｄｉ；②用ＣＤ４０激发型单抗（克隆５Ｃ１１和２Ｇ１１）与ＬＣＤ３２细胞联合ＩＬ－４能使静止期Ｂ细胞体外长期生存和增殖。表明用激发型ＣＤ４０单抗５Ｃ１１和２Ｇ１１建立的ＣＤ４０培养体系，能使Ｂ细胞长期生存和增殖，为体外研究Ｂ细胞提供了有效的手段。

Setup of CD40 System and Longterm Proliferation of B Lymphocytes in Vitro

To establish the CD40 system for longterm culture of human B lymphocytes in vitro by use of antiCD40 monoclonal antibodies prepared in our laboratory,CD32transfected L cells (LCD32) and cytokines.Methods: Flow cytometric analysis for the CD40 molecule expression; Coculture of antiCD40 mAb 5C11 with LCD32 and IL4 as well as other cytokines to set up a CD40 system for observing the viability and proliferation of human tonsil B cells in the CD40 system.Results: CD40 was expressed on B cells from human peripheral blood and tonsil,well expressed by multimyeloma cell line XG2 and B lymphoma cell line Daudi;(2) Agonistic antihuman CD40 mAbs clone 5C11 and 2G11 can combine with LCD32 and IL4 to make resting tonsil B cells survive,form homotypic aggregations and proliferate in vitro in longterm.Thus, the CD40 culture system was successfully established by our agonisti antiCD40 mAbs,providing an efficient way to study the survival as well as proliferation and longterm culturing of human B lymphocytes in vitro