抗人IL-6Rβ(gp130)单克隆抗体对IL-6信号分子调控浅析  

Regulatory effect of anti-IL-6Rβ(gp130)mAb on IL-6 signaling pathway

作　　者：周晓薇[1] 苗平[1] 陈呢喃[1] 赵蓉[1] 钱柳[1] 余奇文[1] 张继英[1] 许荣[1,2] 何东仪 肖连波[1,2] 陆梅生 张冬青 

机构地区：[1]上海交通大学医学院,上海市免疫学研究所,上海200025 [2]上海市长宁区光华中西医结合医院,上海20052

出　　处：《中国免疫学杂志》2014年第5期639-643,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金(No.31270963);上海市科委自然基金(11ZR1427000)资助

摘　　要：目的:分析鼠抗人IL-6Rβ(gp130)单克隆抗体[anti-IL-6Rβ(gp130)mAb]的生物学特性及对IL-6信号分子的调节作用。方法:利用Western blot、竞争性ELISA实验、功能表位结合实验,检测anti-IL-6Rβ(gp130)mAb的亲和力、效价及与gp130抗原表位结合特异性。进而探讨anti-IL-6Rβ(gp130)mAb对U266细胞、RA患者外周血单个核细胞(PBMC)及滑膜液单个核细胞(SFMC)增殖和分化中的IL-6信号分子的调控机制。结果:Anti-IL-6Rβ(gp130)mAb对gp130具有高亲和性(如10A1细胞株的亲和力常数可达为2.62E-10),所获得的多株单抗还具有针对不同抗原结合表位的特性,可用于分析和研究IL-6信号通路及下游STAT3的磷酸化。结论:获得针对不同抗原结合表位的anti-IL-6Rβ(gp130)mAb,初步的实验结果表明此类单抗不仅具有调控IL-6信号分子和下游STAT3磷酸化的生物学功能,并为解析临床RA的发病与IL-6信号分子的关联性提供了初步的实验结果。Objective：To analyse the biological function of anti-IL-6Rβ（gp130） monoclonal antibody and its regulatory effect on IL-6 signaling.Methods：Biological characteristics of anti-IL-6Rβ（gp130） mAb were assessed by Western blot analysis, capture ELISA and peptide ELISA .The phosphorylation of STAT 3 was tested by Western blot analysis in IL-6-stimulated U266/RA-FLS/RA-PBMC with or without anti-IL-6Rβ（gp130） mAb treatment.Results：3 strains of mouse anti-human gp130 mAb were with high affini-ty and different binding epitopes , the kaff of 10A1 was 2.62E-10.In U266, RA-PBMC and RA-SFMC, IL-6 signaling highly activated STAT3 which could be inhibited by anti-gp130 mAb.Conclusion： Anti-IL-6Rβ（ gp130 ） mAb might have different binding epitopes and could affect IL-6 stimulated phosphorylation of STAT3, which provides a preliminary experiment for analyse the correlation of IL-6 signaling and RA .

关 键 词：IL-6 GP130 抗gp130单克隆抗体 STAT3 RA 

分 类 号：R392[医药卫生—免疫学]