Regulation of C3 and factor H synthesis of human glomerular mesangial cells by IL-1 and interferon-gamma.
Previous reports have shown production of complement components C4, C2 and factor B by renal tissue. We have shown recently that human proximal tubular epithelial cells (PTEC) synthesize C3 in vitro, and that IL-2 enhances this production. In the present study we demonstrate that human mesangial cells (MC) in culture produce factor H and that supernatants of activated peripheral blood mononuclear cells (T cell growth factor (TCGF)) induce C3 production and enhance factor H synthesis in both a time- and dose-dependent manner. To investigate whether certain defined cytokines from TCGF were responsible for the observed effect, we tested various cytokines for their effect on complement production by MC. It is shown that IL-1 induces C3 synthesis whereas factor H production is up-regulated by IFN-gamma, in both a dose- and time-dependent manner. Antibody blocking experiments revealed that C3 synthesis induced by both TCGF and IL-1 could be blocked with antibodies specific for IL-1, and also that TCGF and IFN-gamma enhanced factor H synthesis could both be blocked with antibodies specific for IFN-gamma. Cycloheximide was able to inhibit C3 and factor H production, suggesting de novo synthesis of the proteins. mRNA-polymerase chain reaction (PCR) analysis revealed mRNA encoding for C3 after stimulation with TCGF and IL-1. Factor H genes are constitutively expressed in cultured mesangial cells and its expression is up-regulated by TCGF and IFN-gamma. Northern blot analysis with specific probes for C3 and factor H revealed bands which support the results obtained by PCR analysis.
van den Dobbelsteen
M E
ME
Department of Nephrology, University Hospital Leiden, The Netherlands.
Verhasselt
V
V
Kaashoek
J G
JG
Timmerman
J J
JJ
Schroeijers
W E
WE
Verweij
C L
CL
van der Woude
F J
FJ
van Es
L A
LA
Daha
M R
MR
D001483:Base Sequence; D002478:Cells, Cultured; D003176:Complement C3 / Q000096:biosynthesis* / Q000235:genetics; D017242:Complement Factor H / Q000096:biosynthesis* / Q000235:genetics; D005920:Glomerular Mesangium / Q000378:metabolism*; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology*; D007375:Interleukin-1 / Q000494:pharmacology*; D007376:Interleukin-2 / Q000494:pharmacology; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000032:analysis
C493495:CFH protein, human; D003176:Complement C3; D007375:Interleukin-1; D007376:Interleukin-2; D012333:RNA, Messenger; D017242:Complement Factor H; D007371:Interferon-gamma