title: Multiplex analysis of pro-inflammatory cytokines in serum of Actinobacillus pleuropneumoniae-infected pigs.
abstract: Porcine pleuropneumonia is a severe respiratory disease caused by Actinobacillus (A.) pleuropneumoniae. The aim of the present study was to analyze serum samples of A. pleuropneumoniae-infected pigs for TNF-α, IL-1β and IL-6 using a cytometric bead array (CBA) 3-plex assay and additionally for IL-6 using ELISA. The CBA 3-plex assay was successfully validated for use in serum. The limits of detection varied between 0.012 and 0.333 ng/mL, and the inter- and inter-assay coefficients of variation were <5% and <10%, respectively. Increased levels were observed for all 3 cytokines following experimental infection with A. pleuropneumoniae. Mean peak concentrations of TNF-α and IL-6 were recorded at 12h and at 10h p.i., respectively. For IL-6, similar concentration-time profiles were observed with CBA and ELISA. It is proposed that this immuno-assay can be applied for the screening of immunomodulatory properties of drugs and vaccine adjuvants in infection, inflammation and vaccination.
authors.affiliation: Department of Pharmacology, Toxicology and Biochemistry, Ghent University, Faculty of Veterinary Medicine, Salisburylaan 133, 9820 Merelbeke, Belgium. Electronic address: Heidi.Wyns@UGent.be.
authors.forename: H
authors.initials: H
authors.lastname: Wyns
authors.affiliation: Department of Pharmacology, Toxicology and Biochemistry, Ghent University, Faculty of Veterinary Medicine, Salisburylaan 133, 9820 Merelbeke, Belgium.
authors.forename: S
authors.initials: S
authors.lastname: Croubels
authors.affiliation: Laboratory of Immunology, Department of Parasitology, Virology and Immunology, Ghent University, Faculty of Veterinary Medicine, Salisburylaan 133, 9820 Merelbeke, Belgium.
authors.forename: M
authors.initials: M
authors.lastname: Vandekerckhove
authors.affiliation: Department of Pharmacology, Toxicology and Biochemistry, Ghent University, Faculty of Veterinary Medicine, Salisburylaan 133, 9820 Merelbeke, Belgium.
authors.forename: K
authors.initials: K
authors.lastname: Demeyere
authors.affiliation: Department of Pharmacology, Toxicology and Biochemistry, Ghent University, Faculty of Veterinary Medicine, Salisburylaan 133, 9820 Merelbeke, Belgium.
authors.forename: P
authors.initials: P
authors.lastname: De Backer
authors.affiliation: Laboratory of Immunology, Department of Parasitology, Virology and Immunology, Ghent University, Faculty of Veterinary Medicine, Salisburylaan 133, 9820 Merelbeke, Belgium; Vaccine Design, Department of Biosystems KU Leuven, Kasteelpark Arenberg 30, 3001 Heverlee, Belgium.
authors.forename: B M
authors.initials: BM
authors.lastname: Goddeeris
authors.affiliation: Department of Pharmacology, Toxicology and Biochemistry, Ghent University, Faculty of Veterinary Medicine, Salisburylaan 133, 9820 Merelbeke, Belgium.
authors.forename: E
authors.initials: E
authors.lastname: Meyer
mesh_terms: D000189:Actinobacillus Infections / Q000097:blood / Q000276:immunology / Q000662:veterinary*; D016977:Actinobacillus pleuropneumoniae / Q000276:immunology; D000818:Animals; D016207:Cytokines / Q000097:blood*; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005786:Gene Expression Regulation / Q000276:immunology; D007118:Immunoassay; D007249:Inflammation / Q000662:veterinary; D053583:Interleukin-1beta; D013552:Swine; D013553:Swine Diseases / Q000097:blood* / Q000276:immunology
chemical_list: D016207:Cytokines; D053583:Interleukin-1beta
keywords: Actinobacillus pleuropneumoniae; Cytokines; Cytometric Bead Array; Porcine serum