可溶性HLA-G1抗原肽复合物的体外折叠及其功能研究  

In vitro refolding and functional study of soluble HLA-G1-peptide complex

作　　者：杨敬宁[1] 吴雄文[1] 梁智辉[1] 韩军艳[1] 黄亚非[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

出　　处：《细胞与分子免疫学杂志》2005年第3期265-268,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金重点资助项目(No.39830340)

摘　　要：目的:体外折叠形成可溶性HLA G1抗原肽复合物,并研究其生物学功能。方法:以原核表达的可溶性HLA-G1的重链及轻链β2m,与人工合成的抗原九肽体外进行共折叠复性。将折叠产物通过SephadexG75层析柱纯化,经Westernblot进行构象鉴定。探讨可溶性HLA-G1分子对NK细胞杀伤活性的影响,对混合淋巴细胞培养中T细胞增殖的影响,以及对活化T细胞凋亡的影响。结果:折叠产物可与mAbW6/32特异性结合,在Westernblot的鉴定中呈阳性反应。可溶性HLA-G1可有效地抑制NK细胞的细胞毒作用,抑制混合淋巴细胞培养中T细胞的增殖,促进活化T细胞发生凋亡。结论:体外折叠成功地形成可溶性HLA-G1抗原肽复合物,该复合物具有抑制NK细胞和T细胞的活性。AIM: To form soluble HLA-G1-peptide complex by refolding in vitro, and study its immune function. METHODS: The heavy chain and β2m of sHLA-G1 were expressed as insoluble aggregates in E.coli, and then the two subunits were refolded to form HLA-G1-peptide complex by dilution method in the presence of specific peptide. The refolding product was purified through Sephadex G-75 gel filtration. The purified product was identified by Western blot with mAb W6/32. The function of soluble HLA-G1 was explored from following three aspects, namely, the influences on cytotoxicity of NK cells, on proliferation of T cells in mixed lymphocyte culture and apoptosis of activated T cells. RESULTS: The refolded complex was recognized by mAb W6/32. It effectively inhibited cyotoxicity of NK cells and proliferation of T cells, and induced apoptosis of activated T cells. CONCLUSION: The refolding of soluble HLA-G1-peptide complex has been successfully realized in vitro. The complex can inhibit the functions of NK cells and T cells.

关 键 词：sHLA-G1-抗原肽复合物 折叠复性 细胞毒作用 增殖 凋亡 

分 类 号：R392.11[医药卫生—免疫学]