title: Pig α1-acid glycoprotein: characterization and first description in any species as a negative acute phase protein.
abstract: The serum protein α1-acid glycoprotein (AGP), also known as orosomucoid, is generally described as an archetypical positive acute phase protein. Here, porcine AGP was identified, purified and characterized from pooled pig serum. It was found to circulate as a single chain glycoprotein having an apparent molecular weight of 43 kDa by SDS-PAGE under reducing conditions, of which approximately 17 kDa were accounted for by N-bound oligosaccharides. Those data correspond well with the properties of the protein predicted from the single porcine AGP gene (ORM1, Q29014 (UniProt)), containing 5 putative glycosylation sites. A monoclonal antibody (MAb) was produced and shown to quantitatively and specifically react with all microheterogenous forms of pig AGP as analyzed by 2-D electrophoresis. This MAb was used to develop an immunoassay (ELISA) for quantification of AGP in pig serum samples. The adult serum concentrations of pig AGP were in the range of 1-3 mg/ml in a number of conventional pig breeds while it was lower in Göttingen and Ossabaw minipigs (in the 0.3 to 0.6 mg/ml range) and higher in young (2-5 days old) conventional pigs (mean: 6.6 mg/ml). Surprisingly, pig AGP was found to behave as a negative acute phase protein during a range of experimental infections and aseptic inflammation with significant decreases in serum concentration and in hepatic ORM1 expression during the acute phase response. To our knowledge this is the first description in any species of AGP being a negative acute phase protein.
authors.affiliation: Innate Immunology Group, National Veterinary Institute, Technical University of Denmark, Frederiksberg, Denmark. pmhh@vet.dtu.dk
authors.forename: Peter M H
authors.initials: PM
authors.lastname: Heegaard
authors.forename: Ingrid
authors.initials: I
authors.lastname: Miller
authors.forename: Nanna Skall
authors.initials: NS
authors.lastname: Sorensen
authors.forename: Karen Elisabeth
authors.initials: KE
authors.lastname: Soerensen
authors.forename: Kerstin
authors.initials: K
authors.lastname: Skovgaard
mesh_terms: D000209:Acute-Phase Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D000210:Acute-Phase Reaction / Q000097:blood / Q000235:genetics / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000276:immunology; D001665:Binding Sites / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D015870:Gene Expression; D006031:Glycosylation; D054884:Host-Pathogen Interactions; D007249:Inflammation / Q000097:blood / Q000235:genetics / Q000378:metabolism; D008099:Liver / Q000378:metabolism / Q000473:pathology; D008970:Molecular Weight; D009961:Orosomucoid / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D012016:Reference Values; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013290:Streptococcal Infections / Q000097:blood / Q000662:veterinary / Q000821:virology; D017011:Streptococcus suis / Q000502:physiology; D013552:Swine; D013553:Swine Diseases / Q000097:blood / Q000235:genetics / Q000821:virology; D013556:Swine, Miniature
chemical_list: D000209:Acute-Phase Proteins; D000911:Antibodies, Monoclonal; D009961:Orosomucoid