Sublytic complement injury does not activate NF-kappa B, or induce mitogenesis in rat mesangial cells

Exp Nephrol. 2000 Jul-Oct;8(4-5):291-8. doi: 10.1159/000020681.

Abstract

Sublytic complement injury to glomerular mesangial cells, mediated by the terminal membrane attack complex of complement (C5b-9), is a potential initiating mechanism in IgA nephropathy. Sublytic complement injury has been reported to result in the production of a variety of pro-inflammatory molecules and growth factors, including many regulated by the transcription factor NF-kappa B. To determine the importance of complement injury in the pro-inflammatory signalling which occurs in IgA nephropathy, we investigated NF-kappa B activation following sublytic complement injury to cultured rat glomerular mesangial cells (RMCs). A sublytic dose of rabbit anti-Thy 1.1 (THY) serum and normal human serum was selected based upon flow cytometry, chromium-release assay, and induction of superoxide production. No significant C5b-9-induced NF-kappa B activation was detected by electrophoretic mobility shift assays, luciferase activity of RMCs transfected with a NF-kappa B-driven luciferase reporter construct, nor by Northern blots for the NF-kappa B-responsive mRNA species monocyte chemoattractant protein-1 or I kappa B alpha. Furthermore, measurements of (3)H incorporation following sublytic complement injury showed inhibition of mesangial cell mitogenesis in comparison to the heat-inactivated serum treatment and to THY alone. The results of this study suggest that sublytic complement injury to RMC does not directly activate NF-kappa B nor induce mesangial cell proliferation in mesangial cells. Other mechanisms such as IgA immune complex formation must be required to produce these events in IgA nephropathy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies / pharmacology
  • Chromium / metabolism
  • Complement C9 / immunology
  • Complement Membrane Attack Complex / immunology*
  • Electrophoresis
  • Flow Cytometry
  • Glomerular Mesangium / cytology
  • Glomerular Mesangium / immunology*
  • Glomerular Mesangium / metabolism
  • Glomerulonephritis, IGA / immunology
  • Mitosis*
  • NF-kappa B / metabolism*
  • Rats
  • Rats, Sprague-Dawley
  • Superoxides / metabolism
  • Thy-1 Antigens / immunology

Substances

  • Antibodies
  • Complement C9
  • Complement Membrane Attack Complex
  • NF-kappa B
  • Thy-1 Antigens
  • Chromium
  • Superoxides