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.
出处
《现代免疫学》
CAS
CSCD
北大核心
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)
