Inhibition of complement activation on the surface of cells after incorporation of decay-accelerating factor (DAF) into their membranes.
Decay-accelerating factor (DAF), extracted from the stroma of human erythrocytes, was purified to homogeneity and incorporated into the membrane of sheep red cell complement intermediates, where its functional properties were analyzed. Incorporation of DAF into the cell membranes was temperature dependent, took place on pronase- or trypsin-treated erythrocytes, and did not depend on prior deposition of antibody, C1 or C4. Serum lipoproteins (high and low density) effectively inhibited DAF incorporation, but had no effect on the activity of DAF after its association with the cell membrane. The incorporated DAF could not be removed from the red cell surface by repeated washings in the presence of high salt concentration but was solubilized when the stroma were extracted with 0.1% Nonidet P-40. The presence of DAF in the membrane of EA did not affect the deposition of C1 and C4, but as few as 10(2) DAF molecules per cell profoundly inhibited the assembly of C3 and C5 convertases of both the classical and alternative pathways. The DAF inhibitory effect on EAC14 or EAC43 was not overcome by supplying an excess of C2 or factor B, but the alternative pathway C3 convertase could be assembled in the presence of Ni++, or nonphysiological concentrations of Mg++, which enhances the binding affinity of factor B for C3b. The DAF effect on EAC14 or EAC143 was entirely reversed by treating the cells with specific anti-DAF antibodies, showing that DAF did not alter the structure of C4b or C3b. Taken together, the experimental evidence suggests that DAF interacts directly with membrane-bound C3b or C4b and prevents subsequent uptake of C2 and factor B. DAF can function only within the cell membrane. Indeed, the decay dissociation of the C4b2a enzyme on DAF-containing sheep intermediates was not changed by varying the cell concentration. DAF-treated EA had no influence on the decay of nontreated EAC142 present in the same mixture. Moreover, the inhibitory activity of intact human erythrocytes on C4b2a was not blocked by antibodies to DAF, but was abolished by antibodies to the C3b/C4b receptor (CR1). When incorporated into the membrane of rabbit erythrocytes, human DAF inhibited their lysis by human complement. In conclusion, on the basis of these and previous results, it appears that DAF plays a central role in preventing the amplification of the complement cascade on host cell surfaces.(ABSTRACT TRUNCATED AT 400 WORDS)
D000818:Animals; D002352:Carrier Proteins / Q000494:pharmacology; D003166:Complement Activating Enzymes / Q000037:antagonists & inhibitors*; D003167:Complement Activation / Q000187:drug effects; D050577:Complement C3-C5 Convertases / Q000037:antagonists & inhibitors; D003180:Complement C3b Inactivator Proteins / Q000097:blood* / Q000502:physiology; D003181:Complement C4 / Q000378:metabolism; D015935:Complement C4b; D003169:Complement Inactivator Proteins; D004910:Erythrocyte Membrane / Q000201:enzymology* / Q000378:metabolism; D006023:Glycoproteins; D006801:Humans; D008274:Magnesium / Q000494:pharmacology; D009532:Nickel / Q000494:pharmacology; D011817:Rabbits; D011951:Receptors, Complement / Q000502:physiology; D017463:Receptors, Complement 3b; D012756:Sheep
D002352:Carrier Proteins; D003180:Complement C3b Inactivator Proteins; D003181:Complement C4; D003169:Complement Inactivator Proteins; D006023:Glycoproteins; D011951:Receptors, Complement; D017463:Receptors, Complement 3b; D009532:Nickel; D015935:Complement C4b; D003166:Complement Activating Enzymes; D050577:Complement C3-C5 Convertases; D008274:Magnesium