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 language | English |
|---|---|
| Pages (from-to) | 1-14 |
| Number of pages | 14 |
| Journal | Veterinary Immunology and Immunopathology |
| Volume | 81 |
| Issue number | 1-2 |
| Early online date | 3 Aug 2001 |
| DOIs | |
| Publication status | Published - 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