C5a anaphylatoxin as a product of complement activation up-regulates the complement inhibitory factor H in rat Kupffer cells

Eur J Immunol. 2004 Nov;34(11):3257-66. doi: 10.1002/eji.200324806.

Abstract

The 155-kDa complement regulator factor H (FH) is the predominant soluble regulatory protein of the complement system. It acts as a cofactor for the factor I-mediated conversion of the component C3b to iC3b, competes with factor B for a binding site on C3b and C3(H2O) and promotes the dissociation of the C3bBb complex. The primary site of synthesis is the liver, i.e. FH-specific mRNA and protein were identified in both hepatocytes (HC) and Kupffer cells (KC). Previous studies in rat primary HC and KC had shown that the proinflammatory cytokine IFN-gamma influences the balance between activation and inhibition of the complement system through up-regulation of the inhibitory FH. In this study we show that C5a, as a product of complement activation, stimulates the expression of FH-specific mRNA and protein in KC and thus induces a negative feedback. Quantitative-competitive RT-PCR showed an approximate threefold C5a-induced up-regulation of FH. ELISA analyses revealed a corresponding increase in FH protein in the supernatants of KC. The up-regulation of FH was completely inhibited by the C5a-blocking monoclonal antibody 6-9F. Furthermore, an involvement of LPS and IFN-gamma was excluded, which strongly indicates a direct effect of C5a on the expression of FH in KC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Blocking / immunology
  • Blotting, Northern
  • Complement Activation / immunology*
  • Complement C5a / immunology*
  • Complement Factor H / biosynthesis
  • Complement Factor H / immunology*
  • Enzyme-Linked Immunosorbent Assay
  • Kupffer Cells / immunology
  • Kupffer Cells / metabolism
  • Liver / immunology
  • Male
  • RNA, Messenger / biosynthesis
  • RNA, Messenger / genetics
  • Rats
  • Rats, Wistar
  • Reverse Transcriptase Polymerase Chain Reaction
  • Up-Regulation / immunology
  • alpha-N-Acetylgalactosaminidase / immunology

Substances

  • Antibodies, Blocking
  • RNA, Messenger
  • Complement C5a
  • Complement Factor H
  • alpha-N-Acetylgalactosaminidase