Characterization and functional analysis of goldfish (Carassius auratus L.) interleukin-10.
This is the first report of comprehensive functional analysis of an interleukin-10 in bony fish. Quantitative expression analysis of goldfish IL-10 revealed the greatest mRNA levels in the spleen tissues, peripheral blood leukocytes and granulocytes. The stimulation of cells with recombinant goldfish (rg) TNFα2 significantly reduced IL-10 mRNA levels in granulocytes and monocytes of the goldfish. To functionally assess the goldfish IL-10, we generated a recombinant form of the molecule (rgIL-10). The rgIL-10 substantially reduced the expression of TNFα1, TNFα2, IL-1β1, IL-10, CXCL-8, and NADPH oxidase component, p47(phox) in monocytes activated with heat-killed Aeromonas salmonicida and reduced the expression of IFNγ in A. salmonicida-activated splenocytes. Pre-treatment of monocytes with rgIL-10 resulted in substantial reduction of the ROI response of the A. salmonicida or rgIFNγ-primed monocytes. The rgIL-10 bound to goldfish monocytes and induced phosphorylation and nuclear translocation of Stat3. The rgIL-10 also induced rapid and robust increase in the mRNA levels of the goldfish monocyte SOCS-3. Our results indicate that the function of IL-10 is highly conserved through evolution.
Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada.
Leon
L
Grayfer
Jordan W
JW
Hodgkinson
Steven J
SJ
Hitchen
Miodrag
M
Belosevic
D048409:Aeromonas salmonicida / Q000187:drug effects / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D019295:Computational Biology; D003432:Cross-Linking Reagents / Q000378:metabolism; D015536:Down-Regulation / Q000187:drug effects; D020869:Gene Expression Profiling; D006054:Goldfish / Q000276:immunology*; D006358:Hot Temperature; D007371:Interferon-gamma / Q000235:genetics / Q000276:immunology; D016753:Interleukin-10 / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D050296:Microbial Viability / Q000187:drug effects; D008969:Molecular Sequence Data; D009000:Monocytes / Q000187:drug effects / Q000276:immunology / Q000382:microbiology; D010766:Phosphorylation / Q000187:drug effects; D021381:Protein Transport / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology; D050796:STAT3 Transcription Factor / Q000235:genetics / Q000378:metabolism; D050826:Suppressor of Cytokine Signaling Proteins / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects
D003432:Cross-Linking Reagents; D017382:Reactive Oxygen Species; D011994:Recombinant Proteins; D050796:STAT3 Transcription Factor; D050826:Suppressor of Cytokine Signaling Proteins; D016753:Interleukin-10; D007371:Interferon-gamma