Human platelet-initiated formation and uptake of the C5-9 complex of human complement.
We have studied the interaction of radiolabeled complement components with normal human platelets, platelets from a patient with paroxysmal nocturnal hemoglobinuria, and rabbit platelets in the absence of known complement activators or in the presence of cobra venom factor (CVF). When unwashed platelets in platelet-rich plasma, or washed platelets suspended in serum or autologous plasma, were incubated for 30 min, C3 and terminal components (C5, C8, and C9) were found to bind to them. The terminal components were shown to be bound as the C5-9 complex, rather than as individual proteins, by eluting them from the platelet membrane and examining their behavior on ultracentrifugation. They cosedimented at a rate characteristic of the stable C5-9 complex (22S). As many as 370-1,380 C5-9 complexes/platelet were calculated to have been bound during the incubation period. The complex so formed did not differ by ultracentrifugational criteria from that binding to rabbit platelets after CVF activation of complement. Though C3 was not included in the complex, it did not appear to be bound by nonspecific absorption. It could not be removed by washing but rather was eluted by the freeze-thaw technique used to elute the C5-9 complex. Incubation of radiolabeled components in platelet-free plasma did not result in C5-9 complex formation, indicating an initiating role for platelets in this reaction. In contrast to platelets, erythrocytes incubated in analogous plasma did not induce detectable C5-9 formation. Neither EDTA, phenylmethylsulfonylfluoride, nor epsilon-amino-N-caproic acid prevented platelet-initiated formation of C5-9, suggesting that the reaction may involve mechanisms of complement activation not previously described.
D000614:Aminocaproates / Q000494:pharmacology; D000818:Animals; D001666:Binding Sites, Antibody; D001792:Blood Platelets / Q000187:drug effects / Q000276:immunology*; D003182:Complement C5 / Q000096:biosynthesis; D003183:Complement C6 / Q000096:biosynthesis; D003184:Complement C7 / Q000096:biosynthesis; D003185:Complement C8 / Q000096:biosynthesis; D003186:Complement C9 / Q000096:biosynthesis; D003165:Complement System Proteins / Q000096:biosynthesis*; D004492:Edetic Acid / Q000494:pharmacology; D006457:Hemoglobinuria, Paroxysmal / Q000097:blood / Q000276:immunology; D006801:Humans; D066298:In Vitro Techniques; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology; D011817:Rabbits; D012910:Snake Venoms
D000614:Aminocaproates; D003182:Complement C5; D003183:Complement C6; D003184:Complement C7; D003185:Complement C8; D003186:Complement C9; D012910:Snake Venoms; D010664:Phenylmethylsulfonyl Fluoride; D003165:Complement System Proteins; D004492:Edetic Acid