The production and bioactivity of rainbow trout (Oncorhynchus mykiss) recombinant IL-1 beta

S Hong, Jun Zou, M Crampe, S Peddie, Giuseppe Scapigliati, N Bols, C Cunningham, Christopher John Secombes

Research output: Contribution to journalArticle

144 Citations (Scopus)

Abstract

The predicted rainbow trout mature interleukin-1 beta (IL-1 beta) peptide has been produced as a recombinant protein in E. coli, The bioactivity of this molecule has been studied using trout head kidney cell preparations and a trout macrophage cell line (RTS11). Trout rIL-1 beta was shown to increase the expression level of IL-1 beta, cyclooxygenase (COX2) and MHC class II beta chain transcription, as determined by Northern blot analysis. Stimulatory doses of rIL-1 beta were typically greater than or equal to 10 ng/ml. Induction of IL-1 beta expression occurred within 1 h post-stimulation with trout rIL-1 beta and was maximal 3-6 h post-stimulation. Trout rIL-1 beta was also able to increase murine D10.G4.1 cell proliferation and trout head kidney leukocyte phagocytic activity, in a dose-dependent manner. However, equivalent D10.G4.1 cell proliferation was induced with approximately 1000-fold lower doses of human rIL-1 beta. That LPS contamination did not contribute to the effects seen was confirmed by determining its concentration in the trout rIL-1 beta preparation, and demonstrating that the rIL-1 beta activity was inhibited by heating or pre-incubation with a polyclonal anti-trout rIL-1 beta antibody. (C) 2001 Elsevier Science B.V. All rights reserved.

Original languageEnglish
Pages (from-to)1-14
Number of pages14
JournalVeterinary Immunology and Immunopathology
Volume81
Issue number1-2
Early online date3 Aug 2001
DOIs
Publication statusPublished - 30 Aug 2001

Keywords

  • interleukin-1 beta
  • rainbow trout
  • recombinant protein
  • bioactivity testing
  • northern blots
  • interleukin-1 induces interleukin-1
  • molecular-cloning
  • cell-line
  • expression
  • growth
  • gene
  • transcription
  • macrophages
  • protein
  • binding

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