Identification, Molecular Cloning of IL-1β and Its Expression Profile during Nocardia seriolae Infection in Largemouth Bass, Micropterus salmoides

Int J Mol Sci. 2016 Oct 1;17(10):1670. doi: 10.3390/ijms17101670.

Abstract

In the present study, IL-1β cDNA was identified and analyzed from largemouth bass (Micropterus salmoides). Full length IL-1β mRNA was obtained using Rapid Amplification of cDNA Ends (RACE), which contains 78 bp 3'-UTR, a 455 bp 5'-UTR, and an open reading frame (ORF) of 702 bp coding for 233 amino acid residues. The molecular weight and theoretical isoelectric point of largemouth bass IL-1β protein was predicted to be 26.7 kDa and 6.08 respectively. A largemouth bass IL-1β phylogenetic analysis showed a close relation to the IL-1βs of striped trumpeter (Latris lineata), Chinese perch (Siniperca chuatsi), and Japanese sea bass (Lateolabrax japonicus). Peptidoglycan upregulated IL-1β in the spleen and head kidney, while lipopolysaccharide upregulated detectable levels of IL-1β in the spleen only. Largemouth bass, challenged with Nocardia seriolae (1.0 × 10⁶ cfu/mL), showed a significant increase in IL-1β at 3 and 5 days post infection (dpi) in the spleen, while in the head kidney significant expression was found at 2 and 3 dpi, peaking at 3 dpi. Furthermore, tumor necrosis factor α (TNF-α) showed significantly higher expression in the spleen at 3 and 5 dpi, and in the head kidney at 1 and 3 dpi, with expression decreasing at 5 dpi in both tissues.

Keywords: IL-1β; Nocardia seriolae; immunoadjuvant; largemouth bass (Micropterus salmoides).

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Bass / classification
  • Bass / genetics*
  • Bass / immunology*
  • Cloning, Molecular
  • DNA, Complementary / genetics
  • DNA, Complementary / metabolism
  • Fish Diseases / genetics
  • Fish Diseases / immunology*
  • Gene Expression Regulation*
  • Humans
  • Interleukin-1beta / genetics
  • Interleukin-1beta / metabolism*
  • Kidney / metabolism
  • Mice
  • Nocardia / immunology
  • Nocardia Infections / genetics
  • Nocardia Infections / immunology
  • Nocardia Infections / veterinary*
  • Phylogeny
  • Protein Structure, Secondary
  • Rats
  • Sequence Alignment
  • Spleen / metabolism
  • Transcriptome
  • Tumor Necrosis Factor-alpha / genetics
  • Tumor Necrosis Factor-alpha / metabolism

Substances

  • DNA, Complementary
  • Interleukin-1beta
  • Tumor Necrosis Factor-alpha