Functional characterisation of the recombinant tumor necrosis factors in rainbow trout, Oncorhynchus mykiss

Jun Zou, S Peddie, Giuseppe Scapigliati, Y Zhang, N C Bols, A.E. Ellis, Christopher John Secombes

Research output: Contribution to journalArticle

152 Citations (Scopus)

Abstract

Tumor necrosis factor (TNF) is a key mediator in regulating the inflammatory response. Previously two TNF genes have been cloned and sequenced from rainbow trout, Oncorhynchus mykiss. In this study, the mature peptides of the two TNF molecules were produced in bacteria, purified under native conditions and their bioactivities evaluated in vitro. Both trout rTNF1 and rTNF2 induced gene expression of a number of proinflammatory factors including IL1beta, TNF1, TNF2, IL8 and COX2 in freshly isolated head kidney leucocytes and the macrophage cell line RTS11. The stimulatory doses of both rTNFs were greater than or equal to10 ng/ml. Moreover, leucocyte migration and phagocytic activity were enhanced in vitro by the rTNFs in a dose dependent manner. Western blot analysis revealed the presence of multiple forms of rTNF structures including monomeric, dimeric and trimeric forms, suggesting that formation of a homotrimeric structure may be essential for the TNF bioactivities. (C) 2003 Elsevier Science Ltd. All rights reserved.

Original languageEnglish
Pages (from-to)813-822
Number of pages10
JournalDevelopmental and Comparative Immunology
Volume27
Issue number9
Early online date10 Apr 2003
DOIs
Publication statusPublished - Oct 2003

Keywords

  • TNF alpha
  • rainbow trout
  • bioactivity
  • leukocytes in-vitro
  • factor-alpha
  • TNF-alpha
  • molecular-cloning
  • expression
  • receptor
  • cytokines
  • IL-1-beta
  • responses
  • genes

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