人CD40-IgG1Fc段融合蛋白对EBV转化的B细胞凋亡的影响  

Effects of CD40-IgG1Fc fusion protein on apoptosis of EBV-transformed B lymphocytes

作　　者：李和军[1] 张家永[2] 李频[1] 郑祥雄[1] 周小玲[1] 

机构地区：[1]福建医科大学,附属协和医院临床免疫研究所 [2]福建医科大学,闽东医院教学医院风湿科

出　　处：《免疫学杂志》2009年第3期279-282,共4页Immunological Journal

基　　金：福建省科技厅基金资助项目(2003D06)

摘　　要：目的探讨人CD40-IgG1Fc段融合蛋白对EB病毒(EBV)转化的B淋巴细胞在细胞生长及凋亡方面的影响。方法应用流式细胞术检测EBV转化的B淋巴细胞膜表面异位表达CD40L;应用本研究所构建的CD40-IgG1Fc段融合蛋白CHO稳定表达株的培养上清与EBV转化的B淋巴细胞共孵育,通过MTT法检测B细胞的生长变化,吖啶橙(AO)、溴化乙锭(EB)双染色法和DNA ladder法检测融合蛋白对B细胞凋亡的影响。结果EBV转化的B细胞膜表面异位表达CD40L;CD40-IgG1Fc段融合蛋白可有效抑制EB病毒转化的B细胞生长,抑制程度与蛋白浓度呈正相关;AO与EB双染色法及DNA ladder法均检测到B细胞凋亡明显增加,与共孵育时间呈正相关。结论CD40-CD40L的相互作用是EBV转化的B细胞异常激活的重要机制,人CD40-IgG1Fc段融合蛋白能阻断CD40-CD40L的相互作用,抑制EB病毒转化的B细胞的生长,促进其凋亡,为下一步自身免疫性疾病的临床实验治疗奠定了基础。Objective To investigate the effects of CD40-IgG1Fc fusion protein on proliferation and apoptosis of EBV-transformed B lymphocytes. Methods The expression of CD40L on the EBV-transformed B lymphocyte surface was tested by flow cytometry (FCM). After incubation with the fusion protein,the growth of the lymphocyte was tested by MTT colorimetry and the apoptosis was tested by both AO/EB double-staining method and DNA ladder assay. Results CD40L on surface of EBV-transformed B lymphocytes was detected. The growth of the lymphocyte was restrained by CD40-IgG1Fc fusion protein. The increase of apoptosis in the B lymphocyte was detected by both AO/EB double-staining methods and DNA ladder. Conclusion The interaction of CD40-CD40L is an important mechanism for the activation of EBV-transformed B lymphocytes. The fusion protein can obviously restrain the growth and promote the apoptosis of EBV-transformed B lymphocytes by blockage of the interaction of CD40-CD40L. This fusion protein provides a basis for clinical study on treatment of autoimmune diseases.

关 键 词：CD40 IgGlFc段 融合蛋白 B淋巴细胞 凋亡 

分 类 号：R392.12[医药卫生—免疫学]