Identification and expression analysis of two pro-inflammatory cytokines, TNF-α and IL-8, in cobia (Rachycentron canadum L.) in response to Streptococcus dysgalactiae infection

Fish Shellfish Immunol. 2017 Aug:67:159-171. doi: 10.1016/j.fsi.2017.06.014. Epub 2017 Jun 12.

Abstract

Tumor necrosis factor-alpha (TNF-α) and interleukin-8 (IL-8/CXCL8) play pivotal roles in mediating inflammatory responses to invading pathogens. In this study, we identified and analyzed expressions of cobia TNF-α and IL-8 during Streptococcus dysgalactiae infection. The cloned cDNA transcript of cobia TNF-α comprised of 1281 base pairs (bp), with a 774 bp open reading frame (ORF) encoding 257 amino acids. The deduced amino acid sequence of cobia TNF-α showed a close relationship (84% similarity) with TNF-α of yellowtail amberjack. The cloned IL-8 cDNA sequence was 828 bp long, including a 300-bp ORF encoding 99 amino acids. The deduced amino acid sequence of cobia IL-8 shared 90% identity with IL-8 of striped trumpeter. Cobia challenged with a virulent S. dysgalactiae strain displayed an early significant up-regulation of TNF-α and IL-8 in head kidney, liver, and spleen. Notably, IL-8 expression level increased dramatically in the liver at the severe stage of infection (72 h). In conclusion, a better understanding of TNF-α and IL-8 allows more detailed investigation of immune responses in cobia and furthers study on controlling the infectious disease caused by S. dysgalactiae.

Keywords: Cobia; IL-8; Rachycentron canadum; Streptococcus dysgalactiae; TNF-α.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Fish Diseases / immunology*
  • Fish Proteins / chemistry
  • Fish Proteins / genetics*
  • Fish Proteins / immunology
  • Gene Expression Regulation / immunology*
  • Immunity, Innate / genetics*
  • Interleukin-8 / chemistry
  • Interleukin-8 / genetics*
  • Interleukin-8 / immunology
  • Perciformes*
  • Phylogeny
  • Sequence Alignment / veterinary
  • Streptococcal Infections / immunology
  • Streptococcus / physiology
  • Tumor Necrosis Factor-alpha / chemistry
  • Tumor Necrosis Factor-alpha / genetics*
  • Tumor Necrosis Factor-alpha / immunology

Substances

  • Fish Proteins
  • Interleukin-8
  • Tumor Necrosis Factor-alpha