Identification and bioactivities of IFN-gamma in rainbow trout Oncorhynchus mykiss: The First Th1-Type Cytokine Characterized Functionally in Fish

Jun Zou, Allison Christina Carrington, Bertrand Collet, J M Dijkstra, Y Yoshiura, N Bols, Christopher John Secombes

Research output: Contribution to journalArticle

Abstract

IFN-gamma is one of the key cytokines in defining Th1 immune responses. In this study, an IFN-gamma homologue has been identified in rainbow trout Oncorhynchus mykiss, and its biological activities have been characterized. The trout IFN-gamma cDNA is 1034 bp in length and translates into a 180-aa protein. The first intron of the trout IFN-gamma gene contains highly polymorphic GACA minisatellites and 44-bp DNA repeats, giving rise to at least six alleles. IFN-gamma is structurally conserved among vertebrates, and a signature motif has been identified. A nuclear localization sequence known to be crucial for IFN-gamma biological activities is also present in the C-terminal region of the trout IFN-,y. The IFN-gamma expression was induced in head kidney leukocytes by stimulation with PHA or poly(I:Q and in kidney and spleen of fish injected with poly(I:C). rIFN-,y produced in Escherichia coli significantly stimulated gene expression of IFN-gamma-inducible protein 10 (gamma IP-10), MHC class II beta-chain, and STAT1, and enhanced respiratory burst activity in macrophages. Deletion of 29-aa residues from the C terminus containing the nuclear localization sequence motif resulted in loss of activity with respect to induction of gamma IP-10 in RTS-11 cells. Moreover, IFN-gamma-induced gamma IP-10 expression was completely abolished by the protein kinase C inhibitor staurosporine, and partially reduced by U0126, a specific inhibitor for ERKs. Taken together, the present study has demonstrated for the first time a functional IFN-gamma homologue in a fish species, strongly suggesting a conserved Th1 immune response is most likely present in lower vertebrates.

Original languageEnglish
Pages (from-to)2484-2494
Number of pages11
JournalThe Journal of Immunology
Volume175
Issue number4
Publication statusPublished - 15 Aug 2005

Keywords

  • human interferon-GAMMA
  • protein-kinase-C
  • nuclear-localization sequence
  • salmo-gairdneri leukocytes
  • gene
  • expression
  • polymorphism
  • cloning
  • terminus
  • promoter

Cite this

Identification and bioactivities of IFN-gamma in rainbow trout Oncorhynchus mykiss : The First Th1-Type Cytokine Characterized Functionally in Fish. / Zou, Jun; Carrington, Allison Christina; Collet, Bertrand; Dijkstra, J M ; Yoshiura, Y ; Bols, N ; Secombes, Christopher John.

In: The Journal of Immunology, Vol. 175, No. 4, 15.08.2005, p. 2484-2494.

Research output: Contribution to journalArticle

Zou, Jun ; Carrington, Allison Christina ; Collet, Bertrand ; Dijkstra, J M ; Yoshiura, Y ; Bols, N ; Secombes, Christopher John. / Identification and bioactivities of IFN-gamma in rainbow trout Oncorhynchus mykiss : The First Th1-Type Cytokine Characterized Functionally in Fish. In: The Journal of Immunology. 2005 ; Vol. 175, No. 4. pp. 2484-2494.
@article{e709b29b9f18464a990925e1b902b975,
title = "Identification and bioactivities of IFN-gamma in rainbow trout Oncorhynchus mykiss: The First Th1-Type Cytokine Characterized Functionally in Fish",
abstract = "IFN-gamma is one of the key cytokines in defining Th1 immune responses. In this study, an IFN-gamma homologue has been identified in rainbow trout Oncorhynchus mykiss, and its biological activities have been characterized. The trout IFN-gamma cDNA is 1034 bp in length and translates into a 180-aa protein. The first intron of the trout IFN-gamma gene contains highly polymorphic GACA minisatellites and 44-bp DNA repeats, giving rise to at least six alleles. IFN-gamma is structurally conserved among vertebrates, and a signature motif has been identified. A nuclear localization sequence known to be crucial for IFN-gamma biological activities is also present in the C-terminal region of the trout IFN-,y. The IFN-gamma expression was induced in head kidney leukocytes by stimulation with PHA or poly(I:Q and in kidney and spleen of fish injected with poly(I:C). rIFN-,y produced in Escherichia coli significantly stimulated gene expression of IFN-gamma-inducible protein 10 (gamma IP-10), MHC class II beta-chain, and STAT1, and enhanced respiratory burst activity in macrophages. Deletion of 29-aa residues from the C terminus containing the nuclear localization sequence motif resulted in loss of activity with respect to induction of gamma IP-10 in RTS-11 cells. Moreover, IFN-gamma-induced gamma IP-10 expression was completely abolished by the protein kinase C inhibitor staurosporine, and partially reduced by U0126, a specific inhibitor for ERKs. Taken together, the present study has demonstrated for the first time a functional IFN-gamma homologue in a fish species, strongly suggesting a conserved Th1 immune response is most likely present in lower vertebrates.",
keywords = "human interferon-GAMMA, protein-kinase-C, nuclear-localization sequence, salmo-gairdneri leukocytes, gene, expression, polymorphism, cloning, terminus, promoter",
author = "Jun Zou and Carrington, {Allison Christina} and Bertrand Collet and Dijkstra, {J M} and Y Yoshiura and N Bols and Secombes, {Christopher John}",
year = "2005",
month = "8",
day = "15",
language = "English",
volume = "175",
pages = "2484--2494",
journal = "The Journal of Immunology",
issn = "0022-1767",
publisher = "American Association of Immunologists",
number = "4",

}

TY - JOUR

T1 - Identification and bioactivities of IFN-gamma in rainbow trout Oncorhynchus mykiss

T2 - The First Th1-Type Cytokine Characterized Functionally in Fish

AU - Zou, Jun

AU - Carrington, Allison Christina

AU - Collet, Bertrand

AU - Dijkstra, J M

AU - Yoshiura, Y

AU - Bols, N

AU - Secombes, Christopher John

PY - 2005/8/15

Y1 - 2005/8/15

N2 - IFN-gamma is one of the key cytokines in defining Th1 immune responses. In this study, an IFN-gamma homologue has been identified in rainbow trout Oncorhynchus mykiss, and its biological activities have been characterized. The trout IFN-gamma cDNA is 1034 bp in length and translates into a 180-aa protein. The first intron of the trout IFN-gamma gene contains highly polymorphic GACA minisatellites and 44-bp DNA repeats, giving rise to at least six alleles. IFN-gamma is structurally conserved among vertebrates, and a signature motif has been identified. A nuclear localization sequence known to be crucial for IFN-gamma biological activities is also present in the C-terminal region of the trout IFN-,y. The IFN-gamma expression was induced in head kidney leukocytes by stimulation with PHA or poly(I:Q and in kidney and spleen of fish injected with poly(I:C). rIFN-,y produced in Escherichia coli significantly stimulated gene expression of IFN-gamma-inducible protein 10 (gamma IP-10), MHC class II beta-chain, and STAT1, and enhanced respiratory burst activity in macrophages. Deletion of 29-aa residues from the C terminus containing the nuclear localization sequence motif resulted in loss of activity with respect to induction of gamma IP-10 in RTS-11 cells. Moreover, IFN-gamma-induced gamma IP-10 expression was completely abolished by the protein kinase C inhibitor staurosporine, and partially reduced by U0126, a specific inhibitor for ERKs. Taken together, the present study has demonstrated for the first time a functional IFN-gamma homologue in a fish species, strongly suggesting a conserved Th1 immune response is most likely present in lower vertebrates.

AB - IFN-gamma is one of the key cytokines in defining Th1 immune responses. In this study, an IFN-gamma homologue has been identified in rainbow trout Oncorhynchus mykiss, and its biological activities have been characterized. The trout IFN-gamma cDNA is 1034 bp in length and translates into a 180-aa protein. The first intron of the trout IFN-gamma gene contains highly polymorphic GACA minisatellites and 44-bp DNA repeats, giving rise to at least six alleles. IFN-gamma is structurally conserved among vertebrates, and a signature motif has been identified. A nuclear localization sequence known to be crucial for IFN-gamma biological activities is also present in the C-terminal region of the trout IFN-,y. The IFN-gamma expression was induced in head kidney leukocytes by stimulation with PHA or poly(I:Q and in kidney and spleen of fish injected with poly(I:C). rIFN-,y produced in Escherichia coli significantly stimulated gene expression of IFN-gamma-inducible protein 10 (gamma IP-10), MHC class II beta-chain, and STAT1, and enhanced respiratory burst activity in macrophages. Deletion of 29-aa residues from the C terminus containing the nuclear localization sequence motif resulted in loss of activity with respect to induction of gamma IP-10 in RTS-11 cells. Moreover, IFN-gamma-induced gamma IP-10 expression was completely abolished by the protein kinase C inhibitor staurosporine, and partially reduced by U0126, a specific inhibitor for ERKs. Taken together, the present study has demonstrated for the first time a functional IFN-gamma homologue in a fish species, strongly suggesting a conserved Th1 immune response is most likely present in lower vertebrates.

KW - human interferon-GAMMA

KW - protein-kinase-C

KW - nuclear-localization sequence

KW - salmo-gairdneri leukocytes

KW - gene

KW - expression

KW - polymorphism

KW - cloning

KW - terminus

KW - promoter

M3 - Article

VL - 175

SP - 2484

EP - 2494

JO - The Journal of Immunology

JF - The Journal of Immunology

SN - 0022-1767

IS - 4

ER -