Cook D J, Scornik J C
Hum Immunol. 1985 Nov;14(3):234-44. doi: 10.1016/0198-8859(85)90231-9.
HLA class I antigens were purified in sufficient quantities to probe the antigen specificity of lymphocyte and platelet antibodies. Purification of HLA was achieved by affinity chromatography using the monoclonal W6/32 antibody that recognizes a nonpolymorphic determinant of HLA-A,B,C molecules. Pooled platelets from a large number of donors were used as source of antigen. A highly purified HLA preparation was obtained that produced a dose-dependent inhibition of the binding of the W6/32 antibody to lymphocytes as measured by flow cytometry. The binding of monoclonal antibodies to T-lymphocyte antigens or to HLA class II antigens was not affected. The purified HLA also inhibited the binding of lymphocyte alloantibodies from renal transplant patients, providing a direct and definitive way to probe HLA specificity. HLA also inhibited the binding of the same antibodies to platelets but it did not interfere with the binding of alloantibodies to the platelet-specific PLA1 antigen. This preparation, therefore, can conclusively probe the HLA specificity of both alloantibodies for clinical investigation purposes and monoclonal antibodies for screening purposes, and has the potential of becoming a reagent for routine use in clinical and research laboratories.
纯化出足够量的HLA I类抗原，用于探究淋巴细胞和血小板抗体的抗原特异性。通过使用识别HLA - A、B、C分子非多态性决定簇的单克隆W6/32抗体进行亲和层析来实现HLA的纯化。来自大量供体的混合血小板用作抗原来源。获得了一种高度纯化的HLA制剂，通过流式细胞术测量，该制剂对W6/32抗体与淋巴细胞的结合产生剂量依赖性抑制。单克隆抗体与T淋巴细胞抗原或HLA II类抗原的结合不受影响。纯化的HLA还抑制了肾移植患者淋巴细胞同种抗体的结合，为探究HLA特异性提供了一种直接且明确的方法。HLA也抑制了相同抗体与血小板的结合，但不干扰同种抗体与血小板特异性PLA1抗原的结合。因此，该制剂可确凿地探究用于临床研究目的的同种抗体和用于筛选目的的单克隆抗体的HLA特异性，并且有潜力成为临床和研究实验室常规使用的试剂。
