Retinoic acid-inducible gene-I is induced by double-stranded RNA and regulates the expression of CC chemokine ligand (CCL) 5 in human mesangial cells

Nephrol Dial Transplant. 2010 Nov;25(11):3534-9. doi: 10.1093/ndt/gfq270. Epub 2010 May 19.

Abstract

Background: Retinoic acid-inducible gene-I (RIG-I) is a putative RNA helicase involved in immune reactions against RNA viruses and various inflammatory and autoimmune diseases. The purpose of the present study was to investigate the role of RIG-I in glomerular diseases.

Methods: We treated human mesangial cells in culture with polyinosinic-polycytidylic acid (poly IC), which is an authentic double-stranded RNA, and analysed the expression of RIG-I, CC chemokine ligand 5 (CCL5) and interferon (IFN)-β by western blotting, reverse transcriptase-polymerase chain reaction (RT-PCR) or enzyme-linked immunosorbent assay (ELISA). To elucidate the poly IC-signalling pathway, we subjected the cells to RNA interference (RNAi) against RIG-I, IFN-β or Toll-like receptor (TLR) 3. Furthermore, we studied the effects of IFN-β receptor blocking and IFN-β overexpression.

Results: Poly IC induced the expression of RIG-I and CCL5 in human mesangial cells, and RNAi against RIG-I inhibited this poly IC-induced CCL5 expression. Poly IC-induced RIG-I expression was also inhibited by RNAi against IFN-β and by an antibody against the IFN-β receptor. IFN-β overexpression induced the expression of both RIG-I and CCL5. The knockdown of TLR3 abolished poly IC-induced RIG-I expression.

Conclusions: The TLR3/IFN-β/RIG-I/CCL5 signalling pathway may mediate immune and inflammatory responses against viral infection in mesangial cells, suggesting the role of this pathway in the aggravation of glomerulonephritis due to viral infection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cells, Cultured
  • Chemokine CCL5 / genetics*
  • DEAD Box Protein 58
  • DEAD-box RNA Helicases / genetics
  • DEAD-box RNA Helicases / physiology*
  • Dexamethasone / pharmacology
  • Gene Expression Regulation*
  • Humans
  • Interferon-beta / physiology
  • Mesangial Cells / immunology*
  • Poly I-C / pharmacology
  • RNA, Double-Stranded / pharmacology*
  • RNA, Small Interfering / genetics
  • Receptors, Immunologic
  • Toll-Like Receptor 3 / physiology

Substances

  • CCL5 protein, human
  • Chemokine CCL5
  • RNA, Double-Stranded
  • RNA, Small Interfering
  • Receptors, Immunologic
  • TLR3 protein, mouse
  • Toll-Like Receptor 3
  • Interferon-beta
  • Dexamethasone
  • RIGI protein, human
  • DEAD Box Protein 58
  • DEAD-box RNA Helicases
  • Poly I-C