Tanigaki N
Department of Molecular Immunology, Roswell Park Memorial Institute, Buffalo, NY 14263.
Eur J Immunol. 1992 Aug;22(8):2177-80. doi: 10.1002/eji.1830220834.
The specificity and efficiency of endogenous peptides in the HLA class I binding have been investigated by the use of a simple procedure that is based on the serological detection of the folding of HLA class I alpha chains that is induced by the binding to specific peptides in the presence of beta 2 microglobulin. HLA class I proteins were solubilized with a nonionic detergent from cultured HLA homozygous B lymphoblastoid cells and dissociated by alkaline denaturation. The resulting unfolded alpha chains were isolated by gel filtration at a neutral pH. The unfolded alpha chains showed a high refolding capacity and specificity when tested in the presence of an excess beta 2 microglobulin against endogenous peptides extracted by alkaline or acid treatment from cultured B lymphoblastoid cells of various HLA class I phenotypes. Cells of identical or overlapping HLA phenotypes clearly showed shared peptides, whereas such peptide sharing was rarely, if at all, seen between cells of non-overlapping HLA phenotypes. The efficiency of endogenous peptides in the induction of refolding was high; at an estimated concentration of 0.2 microM or less, a strong refolding effect was observed.
通过一种简单的程序研究了内源性肽在HLA I类结合中的特异性和效率，该程序基于对HLA I类α链折叠的血清学检测，这种折叠是在β2微球蛋白存在下与特定肽结合所诱导的。用非离子去污剂从培养的HLA纯合B淋巴母细胞中溶解HLA I类蛋白，并通过碱性变性使其解离。在中性pH下通过凝胶过滤分离得到的未折叠α链。当在过量β2微球蛋白存在下，针对从各种HLA I类表型的培养B淋巴母细胞经碱性或酸性处理提取的内源性肽进行测试时，未折叠的α链显示出高的重折叠能力和特异性。相同或重叠HLA表型的细胞明显显示出共享肽，而非重叠HLA表型的细胞之间很少（如果有的话）见到这种肽共享。内源性肽诱导重折叠的效率很高；在估计浓度为0.2 microM或更低时，观察到强烈的重折叠效应。
