Matsuno K, Diaz-Ricart M, Montgomery R R, Aster R H, Jamieson G A, Tandon N N
Platelet Biology Department, American Red Cross, Rockville, Maryland.
Br J Haematol. 1996 Mar;92(4):960-7. doi: 10.1046/j.1365-2141.1996.422962.x.
Monoclonal anti CD36 antibodies capable of inhibiting platelet adhesion to collagen have not previously been identified. We have now prepared two groups of monoclonal antibodies. One group was prepared using, as immunogen, highly purified (99+%) CD36 prepared by a denaturing procedure. These antibodies (Mo series) reacted strongly with CD36 on protein blots but did not immunoprecipitate native CD36 from platelet lysates nor inhibit platelet adhesion to collagen. The second group of monoclonal antibodies (131 series) was prepared using CD36 purified to >95% by a non-denaturing procedure. These antibodies reacted with control platelets, but not Nak(a)-negative platelets which lack CD36, as measured by flow cytometry and immunoprecipitation. Three monoclonal antibodies of this latter group (131.4, 131.5 and 131.7) inhibited platelet adhesion to collagen in static systems under Mg2+ -independent conditions but had lit tle effect in the presence of Mg2+. 131.4 and 131.7 also inhibited adhesion to collagen using citrated whole blood in a parallel plate flow chamber at physiological shear rates (800s-1), whereas 131.5 was without effect. These are the first anti-CD36 monoclonal antibodies shown to be capable of inhibiting platelet adhesion to collagen and provide further evidence that CD36 plays a role in platelet-collagen interaction.
此前尚未鉴定出能够抑制血小板与胶原蛋白黏附的抗CD36单克隆抗体。我们现已制备了两组单克隆抗体。一组是使用通过变性程序制备的高度纯化（99%以上）的CD36作为免疫原制备的。这些抗体（Mo系列）在蛋白质印迹上与CD36强烈反应，但不能从血小板裂解物中免疫沉淀天然CD36，也不能抑制血小板与胶原蛋白的黏附。第二组单克隆抗体（131系列）是使用通过非变性程序纯化至>95%的CD36制备的。通过流式细胞术和免疫沉淀检测，这些抗体与对照血小板反应，但不与缺乏CD36的Nak(a)阴性血小板反应。后一组中的三种单克隆抗体（131.4、131.5和131.7）在Mg2+非依赖条件下的静态系统中抑制血小板与胶原蛋白的黏附，但在有Mg2+存在时作用很小。131.4和131.7在生理剪切速率（800s-1）下，在平行板流动腔中使用枸橼酸化全血时也抑制与胶原蛋白的黏附，而131.5则无作用。这些是首批被证明能够抑制血小板与胶原蛋白黏附的抗CD36单克隆抗体，并进一步证明CD36在血小板 - 胶原蛋白相互作用中起作用。
