原核表达、体外折叠的HLA-G5促进人外周血单核细胞产生TNF  

The in vitro refolded prokaryotic HLA-G5 promotes the TNF production in human peripheral blood mononuclear cells

作　　者：褚利霞[1] 吴雄文[1] 夏芳珍[1] 鈡茂华 李青[1] 翁秀芳[1] 卢小玲[1] 张彩娥[1] 陆盛军[1] 梁智辉[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

出　　处：《现代免疫学》2006年第3期222-225,共4页Current Immunology

基　　金：国家自然科学基金资助项目(30271201)

摘　　要：通过原核表达获得HLA-G5重链和轻链(β2m),经初步纯化后,利用稀释法与人工合成的九肽(KGPPAALTL)一起进行体外复性折叠,形成HLA-G5-抗原肽复合物,经非变性PAGE/Western blot和夹心ELISA法鉴定,证实复性后成功获得天然构象的HLA-G5。利用PBMC受LPS刺激后产生TNF的特点,观察原核表达、体外折叠的HLA-G5对人PBMC分泌TNF的影响。TNF分泌量采用TNF敏感的L929细胞进行生物法测定,结果显示原核表达、体外折叠的HLA-G5促进人PBMC分泌TNF-α。In this study, the highly expressed and primarily purified prokaryotic HLA-G5 heavy chain（HC）, and the HLA class 1 light ehain（LC, ie. β2m）, refolded in vitro in the presence of a synthetic antigenic peptide （KGPPAALTL）. to form the HLA G5 antigenic peptide complex. The conformation of the refolded complex was detected by HLA class I specific mAb W6/32 with native PAGE/Western blot and ELISA, the effect of the refolded HLA-G5 on the TNF production of human PBMC was observed with bioassay by using TNF-sensitive L929 cell line. The results showed the prokaryotic HLA G5-autigenic peptide complex was successfully produced, sharing the epitope of HLA class 1, and the complex was able, to promote the TNF prochuction of buman PBMC in vitro.

关 键 词：HLA-G5-抗原肽复合物 稀释复性 TNF 

分 类 号：R392.11[医药卫生—免疫学]