Differential regulation of complement factor H and C3 production in human umbilical vein endothelial cells by IFN-gamma and IL-1.
Human umbilical vein endothelial cells (HUVEC) synthesize and secrete C component factors H and C3. Addition of T cell growth factor to HUVEC enhanced factor H production, and caused a profound increase in C3 production. In the present study, investigations were initiated to characterize the effects of purified rIL-1 and rIFN-gamma on the production of factor H and C3 at the protein and mRNA level. IFN-gamma enhanced factor H production in a dose-dependent fashion and a two-fold increase was observed with an optimal dose of 200 U/ml, whereas IFN-gamma had no effect on C3 production. IL-1 inhibited factor H secretion, but the production of C3 was increased 10-fold at an optimal dose of 500 pg/ml of IL-1. Kinetic experiments demonstrated that addition of IL-1 to HUVEC resulted in an induction of C3 production after more than 24 h, whereas IFN-gamma already had a significant effect on factor H production after 8 h of culture. IL-1 in combination with IFN-gamma had a synergistic effect on C3 production. The effects of IL-1 and IFN-gamma on factor H and C3 production by HUVEC could be blocked by using neutralizing amounts of antibodies specific for these cytokines. Northern blot analysis showed that factor H mRNA expression was enhanced in IFN-gamma-treated HUVEC and C3 mRNA was induced in IL-1-treated HUVEC, indicating that the observed increase of factor H and C3 probably is controlled by enhancement of transcription or stability of the transcript.
Brooimans
R A
RA
Department of Nephrology, University Hospital, Leiden, The Netherlands.
van der Ark
A A
AA
Buurman
W A
WA
van Es
L A
LA
Daha
M R
MR
D015152:Blotting, Northern; D002478:Cells, Cultured; D003176:Complement C3 / Q000096:biosynthesis* / Q000235:genetics; D003180:Complement C3b Inactivator Proteins / Q000096:biosynthesis* / Q000235:genetics; D017242:Complement Factor H; D004730:Endothelium, Vascular / Q000378:metabolism*; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D066298:In Vitro Techniques; D007371:Interferon-gamma / Q000494:pharmacology*; D007375:Interleukin-1 / Q000494:pharmacology*; D007376:Interleukin-2 / Q000494:pharmacology; D007700:Kinetics; D012333:RNA, Messenger / Q000235:genetics; D013997:Time Factors
C493495:CFH protein, human; D003176:Complement C3; D003180:Complement C3b Inactivator Proteins; D007375:Interleukin-1; D007376:Interleukin-2; D012333:RNA, Messenger; D017242:Complement Factor H; D007371:Interferon-gamma