Activation of the alternative pathway of complement by cellulosic hemodialysis membranes.
Compared to cellulose acetate, hemodialysis with cuprophan membranes is associated with greater activation of the alternative pathway of complement. Previous studies have shown that this difference is not due to a greater number of potential covalent binding sites for activated C3 on cuprophan. To investigate further the factors that influence complement activation by hemodialysis membranes, proteins were eluted from serum-treated cuprophan and cellulose acetate membranes with hydroxylamine at alkaline pH and analyzed by SDS-PAGE and Western blot. Approximately 23 times more total protein was removed from cellulose acetate. Virtually all the C3 in the cellulose acetate eluate was in the form of inactive fragments C3c and C3dg. In contrast, the functionally active form of C3 (C3b) was a prominent constituent of the cuprophan eluate. The binding of factor B (precursor of the catalytic subunit of the C3 convertase) and factor H (regulatory protein of C3 activation) to serum-treated membranes was also analyzed. By Scatchard's method, the affinity constant at equilibrium for factor B binding (KB) to the two types of membranes was not significantly different; however, there were approximately four times more factor B binding sites on the cuprophan than on the cellulose acetate. For cuprophan, the number of factor B binding sites was 1.6 times greater than the number of factor H binding sites. These studies demonstrate that a portion of the C3b molecules that bind to cuprophan are protected from degradation, and suggest that the complement activating capacity of hemodialysis membranes is determined by biochemical properties that modulate both the binding of serum proteins to the membrane and the interactions of the endogenous regulatory proteins with membrane-associated C3b.
Cheung
A K
AK
Medical Service, Veterans Administration Medical Center, Salt Lake City, Utah.
Parker
C J
CJ
Wilcox
L
L
Janatova
J
J
D002482:Cellulose / Q000031:analogs & derivatives* / Q000494:pharmacology; D003167:Complement Activation / Q000187:drug effects*; D003176:Complement C3 / Q000378:metabolism; D050577:Complement C3-C5 Convertases / Q000378:metabolism; D003180:Complement C3b Inactivator Proteins / Q000378:metabolism; D011415:Complement Factor B / Q000378:metabolism; D017242:Complement Factor H; D003170:Complement Pathway, Alternative / Q000187:drug effects*; D006801:Humans; D007700:Kinetics; D008567:Membranes, Artificial; D008970:Molecular Weight; D010446:Peptide Fragments / Q000302:isolation & purification; D006435:Renal Dialysis
C493495:CFH protein, human; D003176:Complement C3; D003180:Complement C3b Inactivator Proteins; D008567:Membranes, Artificial; D010446:Peptide Fragments; C005062:acetylcellulose; D017242:Complement Factor H; D002482:Cellulose; C009865:cuprammonium cellulose; D050577:Complement C3-C5 Convertases; D011415:Complement Factor B