Cloning and expression analysis of three striped trumpeter (Latris lineata) pro-inflammatory cytokines, TNF-alpha, IL-1beta and IL-8, in response to infection by the ectoparasitic, Chondracanthus goldsmidi

Fish Shellfish Immunol. 2009 May;26(5):773-86. doi: 10.1016/j.fsi.2009.03.012. Epub 2009 Mar 28.

Abstract

This study reports the cloning and sequencing of three striped trumpeter (Latris lineata Forster) pro-inflammatory cytokines, TNF-alpha, IL-1beta and IL-8, as well as their differential expression in response to an infection by the ectoparasite Chondracanthus goldsmidi. The striped trumpeter TNF-alpha transcript consisted of 1093 bp, including a 759 bp ORF which translated into a 253 aa transmembrane peptide. The sequence contained a TACE cut site, that would produce a 167 aa soluble peptide containing the TNF ligand family signature. The IL-1beta sequence consisted of 963 bp, including a 774 bp ORF which translated into a 258 aa protein. The protein lacked both a signal peptide and an ICE cleavage site, but did contain the IL-1 family signature. The sequence for the chemokine IL-8 contained 906 bp, with an ORF of 297 bp, which translated into a 99 aa protein. The protein lacked an ELR motif as is common with many teleost IL-8 sequences. The differential expression of the three cytokine genes in parasitized fish was investigated via quantitative real-time PCR. A significant up-regulation of all three pro-inflammatory cytokines was found in the gills, which were the site of parasite attachment. Examination of head kidney cells revealed a significant up-regulation of TNF-alpha, but not IL-1beta or IL-8. Conversely, the spleen cells showed significant up-regulation of both IL-1beta and IL-8, but not TNF-alpha. These findings allow for more detailed investigations of the striped trumpeter immune response.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cloning, Molecular
  • Copepoda / physiology*
  • Cytokines / chemistry
  • Cytokines / genetics*
  • Ectoparasitic Infestations / immunology
  • Ectoparasitic Infestations / veterinary*
  • Fish Diseases* / immunology
  • Fish Diseases* / parasitology
  • Gene Expression Profiling
  • Gene Expression Regulation / immunology*
  • Gills / cytology
  • Gills / parasitology
  • Interleukin-1beta / genetics
  • Interleukin-8 / genetics
  • Molecular Sequence Data
  • Perciformes / genetics*
  • Perciformes / immunology*
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Sequence Homology, Nucleic Acid
  • Tumor Necrosis Factor-alpha / genetics

Substances

  • Cytokines
  • Interleukin-1beta
  • Interleukin-8
  • Tumor Necrosis Factor-alpha

Associated data

  • GENBANK/FJ532281
  • GENBANK/FJ532282
  • GENBANK/FJ532283