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CD40培养体系的建立及B淋巴细胞的体外培养

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

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【作者】 周照华张学光王江方钱齐宏潘建忠鲁严蒋令瑜

【Author】 Zhou Zhaohua,Zhang Xueguang,Wang Jiangfang,et al (Department of Immunology,Suzhou Medicl College,Suzhou,215007)

【机构】 苏州医学院免疫学研究室

【摘要】 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。

【Abstract】 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

【基金】 国家自然科学基金,青年基金,杰出人才基金
  • 【文献出处】 苏州医学院学报 ,ACTA ACADEMIAE MEDICINAE SUZHOU , 编辑部邮箱 ,1999年07期
  • 【分类号】R392.12
  • 【被引频次】1
  • 【下载频次】267
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