天然免疫分子LILRA1和LILRB2结合并调控HLA—B27的比较性研究  

Comparative study of natural immune molecules LILRA1 and LILRB2 binding and regulating HLA-B27

作　　者：张之勇[1] 任霞[1] 宋冠华[1] 俞林昌 史美艳[1] 郭强[1] 李莲莲[1] 张晓瑜[1] 姜国胜[1] 毕可红[2] 

机构地区：[1]山东省医学科学院基础医学研究所,山东济南250062 [2]山东省千佛山医院,山东济南250014

出　　处：《医学检验与临床》2014年第6期1-5,共5页Medical Laboratory Science and Clinics

基　　金：国家自然科学基金项目（81101605,81172792）;山东省优秀中青年科学家科研奖励基金（BS2009YY008）;山东省自然科学基金（ZR2010HQ034,ZR2009CL014,Y2008C166）;山东省科技发展计划项目（2010GSF10210,2012YD21027,2007GG20002023）;山东省医药卫生科技发展计划项目（2011QZ021）.

摘　　要：目的：探讨天然免疫分子LILRA1，LILRB2与HLA-1类抗原HLA-B27的结合及其调控作用机制。方法：构建LILRA1，LILRB2的真核表达载体和慢病毒载体，利用流式细胞术检测LILRA1，LILRB2在293T细胞系和221-B27细胞系中的表达，利用HLA—B27同源四聚体检测表达在细胞膜表面的LILRA1，LILRB2与HLA—B27分子结合能力，将LILRA1，LILRB2慢病毒载体分别转入HLA—B27稳定转染221细胞后，通过特异性抗体HC10和W6／32检测HLA—B27表达的差异性。结果：测序结果表明LILRA1与LILRB2载体序列正确，LILRA1与LILRB2均能特异性结合HLA—B27同源四聚体，而LILRB2结合HLA—B27能力强于LILRA1；LILRB2稳定转梁的221-B27细胞中HLA—B27分子在细胞膜的表达降低，而L1LRA1的转染对HLA—B27分子在细胞膜的表达无显著性影响。结论：LILRA1，LILRB2均可在细胞膜表面特异的与HLA-B27同源四聚体相结合，LILRB2在细胞内的高表达将抑制HLA—B27分子在细胞膜的表达。Objective ： In order to investigate the binding ability of LILRA1, LILRB2 with HLA-B27, and evaluate the regulation of HLA-B27 by LILRA1 and LILRB2. Methods ： The expression vector pEGFP-LILRA1 and lentivirus vector plenti-LILRA1 were constructed. Both LILRA1 and LILRB2 were transduced into the 293T cells and 221-B27 cells separately, the membrane expression of LILRAI and LILRB2 were detected by flow cytometry, the 293T cells were stained by HLA-B27 homodimertetramers, the expression of HLA-B27 in transduced 221-B27 cells were detected by HC10 and W6/32 antibodies. Results ： The expression vector pEGFP-LILRA1 and plenti-LILRA1 were verified by sequencing; Both LILRA1 and LILRB2 could bind with HLA-B27 homodimer tetramers, and LILRB2 bind with HLA-B27 more strongly than LILRA1; LILRA1 has no effect on HLA-B27 expression in stable LILRA1 transduced 221-B27 cells, while HLA-B27 expression was distinctively inhibited in the LILRB2 stable transduced 221-B27 cells. Conclusions ： Both LILRA1 and LILRB2 could bind with HLA-B27 homodimer tetramer in the cells membrane, and LILRB2 inhibited the expression of HLA-B27.

关 键 词：LILRA1 LILRB2 HLA—B27 慢病毒 

分 类 号：R593.23[医药卫生—内科学]