[In vitro refolding and functional study of soluble HLA-G1-peptide complex].
OBJECTIVE
To form soluble HLA-G1-peptide complex by refolding in vitro, and to study its immune function.


METHODS
The heavy chain and beta(2m) 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 refolded 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 cytotoxicity of NK cells and proliferation of T cells, and induced apoptosis of activated T cells.


CONCLUSIONS
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.
Yang
Jing-ning
JN
Department of Immunology, Tongji Medical College, Huazhong Science and Technology University, Wuhan 430030, China.
Wu
Xiong-wen
XW
Liang
Zhi-hui
ZH
Han
Jun-yan
JY
Huang
Ya-fei
YF
Gong
Fei-li
FL
D000818:Animals; D017209:Apoptosis / Q000276:immunology; D015153:Blotting, Western; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology; D049109:Cell Proliferation; D006680:HLA Antigens / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*; D059951:HLA-G Antigens; D015395:Histocompatibility Antigens Class I / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*; D006801:Humans; D007694:Killer Cells, Natural / Q000276:immunology; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism*; D017510:Protein Folding / Q000187:drug effects; D020673:Protein Renaturation / Q000187:drug effects; D014508:Urea / Q000494:pharmacology
D006680:HLA Antigens; D059951:HLA-G Antigens; D015395:Histocompatibility Antigens Class I; D010455:Peptides; D014508:Urea