Characterization of serum amyloid A (SAA) in rainbow trout using a new monoclonal antibody

Fish Shellfish Immunol. 2014 Oct;40(2):648-58. doi: 10.1016/j.fsi.2014.08.018. Epub 2014 Aug 19.

Abstract

Serum amyloid A (SAA) is an integral part of the innate immune response in mammals and considered to be important during the acute phase response. The present study was undertaken to elucidate the role of SAA protein in the innate immune response of rainbow trout. A monoclonal antibody raised against a recombinant peptide of rainbow trout SAA was characterized using Western blot, dot blot, ELISA and immunohistochemistry. SAA association with high density lipoprotein (HDL) complicated band identification in Western blot, but delipidization of the SAA-HDL isolate highly increased the quality of reaction in the western blot. Rainbow trout fry (87 days post hatch) infected with Yersinia ruckeri showed a significant up-regulation of the SAA gene at 72 h post infection with an increase until 96 h post infection. Non-significant up-regulations were seen at earlier time points i.e. 4 and 24 h. The expression pattern of SAA significantly correlated to the immunohistochemical analysis of the infected fry. A weak staining was seen in liver tissue at 4 h post infection which increased in intensity during the course of infection i.e. 24, 72 and 96 h post infection.

Keywords: Acute phase protein; Innate immune response; Monoclonal antibody; Rainbow trout; SAA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal
  • Fish Diseases / immunology*
  • Fish Diseases / microbiology
  • Fish Proteins / chemistry
  • Fish Proteins / genetics*
  • Fish Proteins / metabolism
  • Immunity, Innate*
  • Oncorhynchus mykiss / genetics*
  • Oncorhynchus mykiss / immunology*
  • Oncorhynchus mykiss / metabolism
  • Real-Time Polymerase Chain Reaction / veterinary
  • Serum Amyloid A Protein / chemistry
  • Serum Amyloid A Protein / genetics*
  • Serum Amyloid A Protein / metabolism
  • Up-Regulation
  • Yersinia Infections / immunology
  • Yersinia Infections / microbiology
  • Yersinia Infections / veterinary*
  • Yersinia ruckeri / physiology

Substances

  • Antibodies, Monoclonal
  • Fish Proteins
  • Serum Amyloid A Protein