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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。
OBJECTIVE: To synthesize the soluble HLA-G1 antigen peptide complex in vitro and study its biological function. Methods: The colifold refolding of soluble HLA-G1 heavy chain and light chain β2m prokaryotic expression with synthetic nonapeptide in vitro. The folded product was purified by Sephadex G75 column and confirmed by Western blot. To investigate the effects of soluble HLA-G1 molecules on cytotoxic activity of NK cells, proliferation of T lymphocytes in mixed lymphocyte culture and apoptosis of activated T cells. Results: The folded product could specifically bind with mAb W6 / 32 and was positive in the identification of Western blot. Soluble HLA-G1 can effectively inhibit the cytotoxicity of NK cells, inhibit the proliferation of T lymphocytes in mixed lymphocyte culture and promote the apoptosis of activated T cells. CONCLUSIONS: In vitro folding successfully formed a soluble HLA-G1 antigen peptide complex, which has the activity of inhibiting NK and T cells.