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为了确定氯奎是否能特异地洗脱血小板表面的HLA抗原和B_2-微球蛋白(β_2-M),作者用4个容积20%(重量/容积)的二磷酸氯奎与1个容积的洗涤血小板(3×10~9/ml)混合,25 C孵育60分钟。再用血小板表面抗原的定量免疫荧光流动细胞计数法和抗血小板膜糖蛋白单克隆抗体检测氯奎处理的血小板,发现70%的HLA抗原和β_2-M从血小板上被洗脱,膜糖蛋白Ⅱb和Ⅲa不受影响。经电泳~(125)I标记的血小板放射自显影,发现除β_2-M外,其它~(125)I标记的膜蛋白不被洗脱。虽然~(125)I不易标记HLA抗原而不能用放射自显影检测,但经抗-HLA单克隆抗体和免疫印迹技术在洗脱液中检测到HLA抗原。经透射电镜检查,氯奎处理的血小板的膜是完整的。这些氯奎处理过的血小板再同血浆孵育,HLA抗原和β_2 M可重
In order to determine whether chloroquine can specifically elute HLA antigens and B_2-microglobulin (β_2-M) on platelet surface, the authors used 4 volumes of 20% (w / v) chloroquine diphosphate and 1 volume of wash Platelets (3 × 10 ~ 9 / ml) were mixed and incubated at 25 ° C for 60 minutes. Then using platelet surface antigen quantitative immunofluorescence flow cytometry and anti-platelet glycoprotein monoclonal antibody detection of chloroquine platelet and found that 70% of HLA antigens and β_2-M was eluted from platelets, membrane glycoprotein Ⅱ b And IIIa are not affected. Electrophoresis ~ (125) I labeled platelet autoradiography, in addition to β_2-M, other ~ (125) I-labeled membrane protein was not eluted. Although ~ (125) I was not easily labeled with HLA antigen and could not be detected by autoradiography, HLA antigens were detected in the eluate by anti-HLA monoclonal antibody and immunoblotting. By transmission electron microscopy, chloroquine-treated platelets are intact. These chloroquine-treated platelets were incubated with plasma again, and HLA antigens and β_2 M were reproducible