Mechanistic studies of the effects of anti-factor H antibodies on complement-mediated lysis.
We have recently reported that complement factor H, a negative regulator of complement-mediated cytotoxicity, is produced and secreted by most bladder cancers. This observation was exploited in the development of the BTA stat and BTA TRAK diagnostic assays, both of which make use of two factor H-specific monoclonal antibodies in sandwich format. Here we show that both antibodies exert interesting effects on the biochemistry of complement activation in in vitro systems. Antibody X13.2 competes with C3b for association with factor H and strongly inhibits factor H/factor I-mediated cleavage of C3b, thereby evidently inactivating a negative regulator of complement; yet, the antibody strongly inhibits complement-mediated lysis as well. Conversely, antibody X52. 1, which does not compete with C3b and has no effect on solution-phase cleavage of C3b, is capable of enhancing complement-mediated lysis of various cell types, including cancer cells, by over 10-fold. Our observations indicate that it is possible to deconvolute the biochemical roles of factor H in complement by means of appropriate inhibitors, a finding with potentially valuable implications for both basic research and cancer therapy.
Corey
M J
MJ
Bion Diagnostic Sciences, Redmond, Washington 98052, USA. michael_corey@biondiagnostics.com
Kinders
R J
RJ
Poduje
C M
CM
Bruce
C L
CL
Rowley
H
H
Brown
L G
LG
Hass
G M
GM
Vessella
R L
RL
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D015153:Blotting, Western; D003167:Complement Activation / Q000187:drug effects*; D003179:Complement C3b / Q000378:metabolism; D017242:Complement Factor H / Q000276:immunology*; D004305:Dose-Response Relationship, Drug; D004912:Erythrocytes / Q000276:immunology; D018922:HL-60 Cells; D006461:Hemolysis / Q000276:immunology; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007700:Kinetics; D008954:Models, Biological; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012756:Sheep; D013997:Time Factors; D014407:Tumor Cells, Cultured; D015854:Up-Regulation; D015054:Zymosan / Q000378:metabolism
D000911:Antibodies, Monoclonal; C493495:CFH protein, human; D003179:Complement C3b; D017242:Complement Factor H; D015054:Zymosan