Biosynthesis of complement factor H by human umbilical vein endothelial cells. Regulation by T cell growth factor and IFN-gamma.
The present studies were initiated to characterize a 150-kDa molecule with inhibitory activity for C3bBb formation, which is present in human umbilical vein endothelial cells (HUVEC). Therefore, human endothelial culture supernatants (HECS) were analyzed for the presence of human complement factor H by ELISA. It was found that H was present in HECS. An immunoblot analysis of affinity purified H from HECS showed that the size of HUVEC H was identical to that of plasma H. The mean production of H by HUVEC of first passage cultures was 40 ng/10(6) cells/day. The synthesis of HUVEC H was fully inhibitable by the addition of cycloheximide to the cultures, suggesting that H is de novo synthesized. Additional evidence for de novo synthesis was obtained by using biosynthetic labeling with [35S] methionine, immunoprecipitation, and SDS-PAGE. It was demonstrated that, indeed, HUVEC produce and secrete factor H. Two forms of the protein were identified, the 150-kDa form and also a 45-kDa form, both forms have been identified in plasma. The functional activity of HUVEC H is identical to that of plasma H. IFN-gamma induced enhanced synthesis of H by HUVEC, whereas it had no effect on C3 synthesis. Supernatant from stimulated PBMC, T cell growth factor, enhanced synthesis of both H and C3. The present studies indicate that H is produced by HU-VEC and that H may function as an inhibitor of complement activation at the endothelial cell level and, thereby, together with molecules like decay accelerating factor and membrane cofactor protein, may influence resistance of endothelial cells to complement mediated damage.
Brooimans
R A
RA
Department of Nephrology, University Hospital Leiden, The Netherlands.
Hiemstra
P S
PS
van der Ark
A A
AA
Sim
R B
RB
van Es
L A
LA
Daha
M R
MR
D001667:Binding, Competitive; D002474:Cell-Free System; D002478:Cells, Cultured; D003179:Complement C3b / Q000096:biosynthesis; D003180:Complement C3b Inactivator Proteins / Q000096:biosynthesis* / Q000302:isolation & purification / Q000502:physiology; D017242:Complement Factor H; D004730:Endothelium, Vascular / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D007371:Interferon-gamma / Q000494:pharmacology*; D007376:Interleukin-2 / Q000494:pharmacology*; D011994:Recombinant Proteins; D014471:Umbilical Veins
C493495:CFH protein, human; D003180:Complement C3b Inactivator Proteins; D007376:Interleukin-2; D011994:Recombinant Proteins; D003179:Complement C3b; D017242:Complement Factor H; D007371:Interferon-gamma