Kao K J
Department of Pathology, University of Florida, Gainesville.
Transfusion. 1988 Jan-Feb;28(1):14-7. doi: 10.1046/j.1537-2995.1988.28188127942.x.
To determine whether chloroquine can specifically elute HLA antigens and beta 2-microglobulin (beta 2-M) from the platelet surface, quantitative immunofluorescence flow cytometry and monoclonal antibodies were used to show that HLA antigens and beta 2-M were proportionally eluted from the platelet surface without affecting the membrane glycoproteins IIb and IIIa. Second, an autoradiogram of electrophoresed I125-labeled platelets showed that only beta 2-M but not other I125-labeled membrane proteins could be eluted. Although HLA antigens were poorly labeled by I125 and could not be detected on the autoradiogram, the eluted HLA antigens could be detected by anti-HLA monoclonal antibody and immunoblotting techniques. No loss of plasma membrane integrity was observed by transmission electron microscopy after chloroquine treatment of platelets. The results indicate that chloroquine selectively elutes HLA antigens and their noncovalently associated beta 2-M without affecting other integral platelet membrane proteins.
为了确定氯喹是否能从血小板表面特异性洗脱HLA抗原和β2-微球蛋白(β2-M),采用定量免疫荧光流式细胞术和单克隆抗体来表明HLA抗原和β2-M从血小板表面按比例洗脱,而不影响膜糖蛋白IIb和IIIa。其次,对经电泳的125I标记血小板的放射自显影片显示,只有β2-M可被洗脱,而其他125I标记的膜蛋白则不能。虽然HLA抗原被125I标记得很差,在放射自显影片上无法检测到,但洗脱的HLA抗原可通过抗HLA单克隆抗体和免疫印迹技术检测到。用氯喹处理血小板后,透射电子显微镜未观察到质膜完整性的丧失。结果表明,氯喹能选择性洗脱HLA抗原及其非共价结合的β2-M,而不影响血小板其他完整的膜蛋白。