Rainbow trout interleukin-2: cloning, expression and bioactivity analysis

P Díaz-Rosales, S Bird, T H Wang, K Fujiki, W S Davidson, J Zou, C J Secombes

Research output: Contribution to journalArticle

56 Citations (Scopus)

Abstract

In this study the rainbow trout (Oncorhynchus mykiss) interleukin-2 (IL-2) cDNA has been cloned, and its expression and bioactivity analysed in head kidney leucocytes. The IL-2 precursor encoded an open reading frame of 429 bp, that translates into a predicted protein of 142 aa, with a 20 aa signal peptide. The trout IL-2 had moderate protein homology (30.9% identity/48.3% similarity) with Fugu IL-2, the only IL-2 homologue identified in fish to date, with lower homology to avian (17.8% identity/23.2% similarity) and mammalian (34.2 identity/46.5% similarity) IL-2s. IL-2 expression was induced by the T cell mitogen PHA and by the mixed leucocyte reaction, where leucocytes from pairs of fish were cultured together for four days. Expression was also induced in vivo during bacterial (Yersinia ruckeri) infection. The Escherichia coli produced recombinant IL-2 was shown to increase the expression of two transcription factors, STAT5 and Blimp-1, known to be involved in IL-2 signalling in mammals, as well as IFN-gamma, gIP and IL-2 itself The potential signalling pathways involved and possible use as an adjuvant for fish vaccines are discussed. (C) 2009 Elsevier Ltd. All rights reserved.

Original languageEnglish
Pages (from-to)414-422
Number of pages9
JournalFish & Shellfish Immunology
Volume27
Issue number3
Early online date21 Jun 2009
DOIs
Publication statusPublished - Sep 2009

Keywords

  • amino acid sequence
  • animals
  • base sequence
  • cloning, molecular
  • fish diseases
  • gene expression profiling
  • gene expression regulation
  • interleukin-2
  • leukocytes
  • lymphoid Tissue
  • molecular sequence data
  • Oncorhynchus mykiss
  • recombinant proteins
  • sequence alignment
  • time factors
  • yersinia infections
  • yersinia ruckeri
  • functional-characterization
  • recombinant IL-2
  • cloning
  • expression
  • STAT5
  • blimp-1
  • IL-1 beta 1
  • IL-2
  • IFN-gamma 1
  • gIP
  • COX-2
  • receptor-gamma-chain
  • oncorhynchus-mykiss walbaum
  • JAK-3 janus kinase
  • cell growth-factor
  • murine-B-lymphoma
  • IL-2 receptor
  • terminal differentiation
  • molecular-cloning
  • t-cells

Cite this

Rainbow trout interleukin-2 : cloning, expression and bioactivity analysis. / Díaz-Rosales, P; Bird, S; Wang, T H; Fujiki, K; Davidson, W S; Zou, J; Secombes, C J.

In: Fish & Shellfish Immunology, Vol. 27, No. 3, 09.2009, p. 414-422.

Research output: Contribution to journalArticle

Díaz-Rosales, P ; Bird, S ; Wang, T H ; Fujiki, K ; Davidson, W S ; Zou, J ; Secombes, C J. / Rainbow trout interleukin-2 : cloning, expression and bioactivity analysis. In: Fish & Shellfish Immunology. 2009 ; Vol. 27, No. 3. pp. 414-422.
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N2 - In this study the rainbow trout (Oncorhynchus mykiss) interleukin-2 (IL-2) cDNA has been cloned, and its expression and bioactivity analysed in head kidney leucocytes. The IL-2 precursor encoded an open reading frame of 429 bp, that translates into a predicted protein of 142 aa, with a 20 aa signal peptide. The trout IL-2 had moderate protein homology (30.9% identity/48.3% similarity) with Fugu IL-2, the only IL-2 homologue identified in fish to date, with lower homology to avian (17.8% identity/23.2% similarity) and mammalian (34.2 identity/46.5% similarity) IL-2s. IL-2 expression was induced by the T cell mitogen PHA and by the mixed leucocyte reaction, where leucocytes from pairs of fish were cultured together for four days. Expression was also induced in vivo during bacterial (Yersinia ruckeri) infection. The Escherichia coli produced recombinant IL-2 was shown to increase the expression of two transcription factors, STAT5 and Blimp-1, known to be involved in IL-2 signalling in mammals, as well as IFN-gamma, gIP and IL-2 itself The potential signalling pathways involved and possible use as an adjuvant for fish vaccines are discussed. (C) 2009 Elsevier Ltd. All rights reserved.

AB - In this study the rainbow trout (Oncorhynchus mykiss) interleukin-2 (IL-2) cDNA has been cloned, and its expression and bioactivity analysed in head kidney leucocytes. The IL-2 precursor encoded an open reading frame of 429 bp, that translates into a predicted protein of 142 aa, with a 20 aa signal peptide. The trout IL-2 had moderate protein homology (30.9% identity/48.3% similarity) with Fugu IL-2, the only IL-2 homologue identified in fish to date, with lower homology to avian (17.8% identity/23.2% similarity) and mammalian (34.2 identity/46.5% similarity) IL-2s. IL-2 expression was induced by the T cell mitogen PHA and by the mixed leucocyte reaction, where leucocytes from pairs of fish were cultured together for four days. Expression was also induced in vivo during bacterial (Yersinia ruckeri) infection. The Escherichia coli produced recombinant IL-2 was shown to increase the expression of two transcription factors, STAT5 and Blimp-1, known to be involved in IL-2 signalling in mammals, as well as IFN-gamma, gIP and IL-2 itself The potential signalling pathways involved and possible use as an adjuvant for fish vaccines are discussed. (C) 2009 Elsevier Ltd. All rights reserved.

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KW - lymphoid Tissue

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KW - recombinant proteins

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KW - receptor-gamma-chain

KW - oncorhynchus-mykiss walbaum

KW - JAK-3 janus kinase

KW - cell growth-factor

KW - murine-B-lymphoma

KW - IL-2 receptor

KW - terminal differentiation

KW - molecular-cloning

KW - t-cells

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JO - Fish & Shellfish Immunology

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