[In vitro refolding and functional study of soluble HLA-G1-peptide complex].
作者信息
Yang Jing-ning, Wu Xiong-wen, Liang Zhi-hui, Han Jun-yan, Huang Ya-fei, Gong Fei-li
机构信息
Department of Immunology, Tongji Medical College, Huazhong Science and Technology University, Wuhan 430030, China.
出版信息
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2005 May;21(3):265-8.
AIM
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.
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.
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