Effects of various types of anti-human platelet membrane antibody on platelet function.
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概要
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Antibodies I and II (heterologous anti-human platelet membrane antisera) were produced in rabbit against human platelet membrane dissolved in 10mM Tris/HCl buffer, pH 7.4, containing 1% Triton X-100, 0.25M sucrose, 2mM CaCl2, and in 38mM Tris/HCl buffer, pH 8.6, containing 1% Triton X-100, 0.1M glycine, 5mM EDTA, respectively. Antibody III (mouse monoclonal antibody) was raised against crude human platelet membrane of normal platelets. By crossed immunoelectrophoresis (CIE), it was shown that antibody I could bind to a single protein. Since the immunoprecipitate on CIE was not detectable by using platelets from a Glanzmann's thrombasthenic patient, antibody I was considered to be specific antibody against GP IIb/IIIa complex. Avidin biotin complex system (ABC)-immunoblot showed the strong binding of antibody II to electrophoretically separated GP IIb and GP IIIa. ABC-immunoblot revealed no binding of antibody III to platelet proteins after SDS-PAGE, but two immunoprecipitates were obtained with an apparent molecular weight of 130K and 110K under reduced conditions by double antibody immunoprecipitate, thus indicating that antibody III was a monoclonal antibody against GP IIb/IIIa complex. Fluorescence flow cytometric analysis showed that the activity of antibody I to sensitize human platelets was less than that of antibody II. In contrast, antibody I was more potent in inhibiting ADP-and collagen-induced platelet aggregation than antibody II. Antibody III inhibited ADP-and collagen-induced aggregation in a dose dependent manner. The addition of 6.3μg/ml of antibody III to platelets sensitized 68% of platelets, and caused complete inhibition of ADP-induced aggregation, while its addition of 3.1μg/ml sensitized 50% of platelets, and caused complete inhibition of collagen-induced aggregation. The results give further support to the proposal that platelet GP IIb/IIIa complex, not dissociated GP IIb and GP IIIa, plays an important role in platelet aggregation.
- 一般社団法人 日本血栓止血学会の論文
一般社団法人 日本血栓止血学会 | 論文
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