原核表达、稀释复性的可溶性HLA-G1抑制混合淋巴细胞培养中T细胞增殖  

The prokaryotic sHLA-G1 refolded in vitro inhibits T cell proliferation in mixed lymphocytes culture

作　　者：夏芳珍[1,2] 钟茂华[1,2] 张彩娥[1,2] 陆盛军[1,2] 吴雄文[1,2] 梁智辉[1,2] 龚非力[1,2] 

机构地区：[1]华中科技大学同济医学院免疫教研室,武汉430030 [2]上海市第九人民医院内分泌实验室,上海200011

出　　处：《中华微生物学和免疫学杂志》2007年第4期299-301,共3页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助（No.30271201）;科技部973计划资助课题（No.2001CB510008）

摘　　要：目的：探讨原核表达体外稀释复性的可溶性人类白细胞抗原G1（soluble　HLA-G1，sHLA-G1）对体外混合淋巴细胞反应中T细胞增殖的影响。方法利用原核表达的sHIA-G1的重链、轻链及人工合成的九肽，折叠形成正确构象的sHLA-G1．肽复合物；在混合淋巴细胞培养（MLC）体系中加入折叠所得的sHLA-G1-肽复合物及相同浓度的对照物，流式细胞仪检测T细胞的增殖情况。结果sHLA-G1．肽复合物加入到混合淋巴细胞培养中能够抑制T细胞的增殖，3个个体的抑制率分别为28．5％、32．5％及22．1％（Dunnett t检验，P〈0．05）；这种抑制作用可以被HLA-G特异性McAb（G11E5）所逆转。结论通过原核表达，体外稀释复性的方法可以大量制备正确构象的sHLA-G1-肽复合物，该复合物分子能够抑制混合淋巴细胞培养中T细胞的增殖。Objective To study the effect of soluble human leukocyte antigen G1 （sHLA-G1） prepared by refolding in vitro on the proliferation of T cells in mixed lymphocyte reaction （MLR）.Methods The sHLA- Gl-peptide complex was formed by refolding in vitro with prokaryotic sHLA-G1 heavy chain, β2 M and a synthesized, HLA-G restricted nonapeptide. Peripheral blood lymphocytes（PBL） of three healthy donors were labeled with CFSE, and co-cultured with lymphoblastoid cells line E001 inactivated by Mitomycine C.Three hundred μg/ml sHLA-G1 and the same concentration of different controls were added into the co-culture. Results The proliferation indexes for the PBL of the three donors against the allogeneic E001 are 7.3500±0.4879,8.9700±0.2353 and 7.4600±0.5726, which are significantly different （5.5266±0.6124, 6.0566±0.4660 and 5.8133±0.3470, respectively） when the co-culture are treated with the sHLA-Gl-peptide complex. The proliferation-in-hibiting effect can be blocked by HLA-G specific monoclonal antibody GllE5. Conchusion Prokaryotic sHLA-G1 refolded in vitro inhibits the proliferation of responder T cells in MLC and the inhibition can be blocked by the HLA-G-specific monoclonal antibody GllE5.

关 键 词：T细胞增殖 人类白细胞抗原G 折叠复性 混合淋巴细胞反应 

分 类 号：R686[医药卫生—骨科学]