Multiplex analysis of pro-inflammatory cytokines in serum of Actinobacillus pleuropneumoniae-infected pigs

Res Vet Sci. 2015 Oct:102:45-8. doi: 10.1016/j.rvsc.2015.07.006. Epub 2015 Jul 14.

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.

Keywords: Actinobacillus pleuropneumoniae; Cytokines; Cytometric Bead Array; Porcine serum.

MeSH terms

  • Actinobacillus Infections / blood
  • Actinobacillus Infections / immunology
  • Actinobacillus Infections / veterinary*
  • Actinobacillus pleuropneumoniae / immunology
  • Animals
  • Cytokines / blood*
  • Enzyme-Linked Immunosorbent Assay / veterinary
  • Gene Expression Regulation / immunology
  • Immunoassay
  • Inflammation / veterinary
  • Interleukin-1beta
  • Swine
  • Swine Diseases / blood*
  • Swine Diseases / immunology

Substances

  • Cytokines
  • Interleukin-1beta