可溶性小鼠BTLA对树突状细胞表面B7-1和B7-2表达的影响  

Effect of murine soluble BTLA on the expression of B7-1 and B7-2 on dendritic cells

作　　者：徐军发[1] 赵坤[2] 黄保军[1] 熊平[1] 冯玮[1] 徐勇[1] 方敏[1] 郑芳[1] 龚非力[1] 

机构地区：[1]华中科技大学同济医学院免疫学系,武汉430030 [2]广东医学院临床免疫学教研室,湛江524023

出　　处：《中华微生物学和免疫学杂志》2006年第12期1032-1037,共6页Chinese Journal of Microbiology and Immunology

基　　金：国家重点基础研究发展规划(973)资助(No.2001cb500008);教育部博士点基金资助(No.20040487056)

摘　　要：目的研究可溶性小鼠B／T淋巴细胞弱化因子(murine B and T lymphocyte attenuator, mBTLA)-人IgG1 Fc融合蛋白(mBTLA-hIg)对树突状细胞表面共刺激分子表达的影响。方法克隆mBTIA基因,构建mBTLA胞外功能区和人IgG1 Fc融合基因的真核表达载体(pmBTLA-hIg),并采用脂质体转染法,将pmBTLA-hIg质粒转染小鼠树突状细胞系(DC2.4)。采用RT-PCR、ELISA和Western blot分别检测pmBTLA-hIg质粒转染DC中mBTLA基因mRNA的表达及细胞培养上清中mBTLA-hIg融合蛋白的表达;采用流式细胞术检测pmBTLA-hIg质粒转染对DC2．4表面共刺激分子B7-1(CD80)和B7-2 (CD86)表达的影响。结果成功克隆了小鼠BTLA基因,并构建了真核表达载体;mBTLA-hIg融合基因转染的DC高表达mBTLA-hIg融合蛋白,并可结合到DC表面。表达可溶性mBTLA-hIg融合蛋白的DC2．4表面B7-1的表达上调,B7-2的表达下调,而且这种改变可被兔抗GST-mBTLA血清阻断。结论可溶性mBTLA-hIg融合蛋白对DC细胞表面B7分子的表达具有调节作用,可能是BTLA反向信号作用于DC的结果。Objective To study the effect of soluble murine B and T lymphocyte attemuator （mBTLA） and human IgG1 Fc fusion protein （mBTLA-hIg） on the expression of B7-1 and B7-2 on dendritic cells （DC）. Methods An eukaryotic expression vector （pmBTLA-hIg）, which expressed soluble mBTLA-hIg fusion protein, was constructed. Then the plasmid was transfected into a mouse dendritic cell line. DC2.4, origining from C57BL/6（B6） mouse, by LipofectAMINE^TM 2000. The expression of mBTLA mRNA and mBTLA-hIg fusion protein were determined by RT-PCR, ELISA and Western blot. The effect of mBTLA-hIg fusion protein on the expression of costimulatory molecules, B7-1 （CD80） and B7-2 （CD86）, on DC was analyzed by flow cytometry. Results The eukaryotic expression vector（pmBTLA-hIg） was constructed and confirmed by restriction endonucleases digestion and DNA sequencing. pmBTLA-hIg transfected DC2.4 could express mBTLA mRNA and release a 43.3×10^3 fusion protein. The fusion protein could binding to DC, and then up-regulated the expression of B7-1 （CD80） , but down-regulated the expression of B7-2 （CD86）, on DC2.4. Moreover the change of the expression of B7-1 and B7-2 could block by the addition of anti-GST-mBTLA serum. Conclusion Soluble mBTLA-hIg fusion protein can regulate the co-stimulatory molecules expression, which might be resulted from the reverse signal of BTLA.

关 键 词：树突状细胞 共刺激分子 BIT淋巴细胞弱化因子 疱疹病毒进入介质 

分 类 号：R392[医药卫生—免疫学]