Cloning and expression analysis of two pro-inflammatory cytokines, IL-1beta and IL-8, in haddock (Melanogrammus aeglefinus).

Yolanda Corripio Miyar, Steven Bird, Konstantinos Tsamopoulos, Christopher John Secombes

Research output: Contribution to journalArticle

111 Citations (Scopus)

Abstract

This paper reports the cloning and sequencing of two pro-inflammatory cytokines, interleukin (IL)-1 beta and IL-8, in haddock (Melanogrammus aeglefinus) by homology cloning. The complete transcript of the haddock IL-1 beta was sequenced and contained 1043 bp, including a 762 bp open reading frame. The 3' end of the gene includes a polyadenylation signal 13 bp upstream of the poly(A) tail, along with 10 instability motifs. The predicted protein of 253aa revealed the presence of the IL-1 family signature and the absence of an ICE cut site. The cDNA of the chemokine IL-8 was sequenced in haddock and contained 903 bp of which 306 bp are the open reading frame. Interestingly, the predicted protein sequence of 101aa, contains an ELR motif preceding the CXC signature, common in all vertebrate IL-8 molecules but absent in all teleost genes sequenced to date. The expression of both haddock cytokines was studied in four different tissues: head kidney, spleen, liver and gill. Tissues were obtained from both healthy fish and fish stimulated in vivo with four commercial serotypes of LPS, namely Escherichia coli 026:136, 055:135, 0111:134 and 0 127:138 and PMA. Haddock IL-1 beta was not constitutively expressed and expression was only observed following stimulation. However, this expression was stimulant dependent and only PMA and LPS 026:136 induced high levels of expression in the head kidney. The haddock IL-8 gene on the other hand, showed a constitutive expression, that could be up or down-regulated depending on the immunostimulant used, although to a lesser extent than IL-1 beta. (c) 2006 Elsevier Ltd. All rights reserved.

Original languageEnglish
Pages (from-to)1361-1373
Number of pages13
JournalMolecular Immunology
Volume44
Issue number6
Early online date10 Jul 2006
DOIs
Publication statusPublished - Feb 2007

Keywords

  • haddock
  • cytokines
  • interleukin
  • LPS serotypes
  • ELR motif
  • trout oncorhynchus-mykiss
  • Gadus-Morhua L.
  • humoral immune parameters
  • molecular-cloning
  • Atlantic Cod
  • larval haddock
  • different serotypes
  • sequence alignment
  • in-vivo
  • interleukin-1-beta

Cite this