title: Characterization of serum amyloid A (SAA) in rainbow trout using a new monoclonal antibody.
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.
authors.affiliation: Laboratory of Aquatic Pathobiology, Department of Veterinary Disease Biology, University of Copenhagen, Stigbojlen 7, DK-1870 Frederiksberg, Denmark. Electronic address: pwk@sund.ku.dk.
authors.forename: Per W
authors.initials: PW
authors.lastname: Kania
authors.affiliation: Laboratory of Aquatic Pathobiology, Department of Veterinary Disease Biology, University of Copenhagen, Stigbojlen 7, DK-1870 Frederiksberg, Denmark.
authors.forename: Jiwan K
authors.initials: JK
authors.lastname: Chettri
authors.affiliation: Laboratory of Aquatic Pathobiology, Department of Veterinary Disease Biology, University of Copenhagen, Stigbojlen 7, DK-1870 Frederiksberg, Denmark.
authors.forename: Kurt
authors.initials: K
authors.lastname: Buchmann
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D005393:Fish Diseases / Q000276:immunology* / Q000382:microbiology; D029941:Fish Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D007113:Immunity, Innate; D017686:Oncorhynchus mykiss / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D000685:Serum Amyloid A Protein / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015854:Up-Regulation; D015009:Yersinia Infections / Q000276:immunology / Q000382:microbiology / Q000662:veterinary*; D044124:Yersinia ruckeri / Q000502:physiology
chemical_list: D000911:Antibodies, Monoclonal; D029941:Fish Proteins; D000685:Serum Amyloid A Protein
keywords: Acute phase protein; Innate immune response; Monoclonal antibody; Rainbow trout; SAA