Detection of chicken interleukin-10 production in intestinal epithelial cells and necrotic enteritis induced by Clostridium perfringens using capture ELISA

Vet Immunol Immunopathol. 2018 Oct:204:52-58. doi: 10.1016/j.vetimm.2018.10.001. Epub 2018 Oct 11.

Abstract

Aims: To assess the production level of interleukin (IL)-10 inClostridium perfringens (CP)-stimulated intestinal epithelial cells (IECs) and CP-infected chickens using an antigen capture ELISA developed by mouse monoclonal antibodies against chicken IL-10. Also, to investigate which CP toxins induced IL-10 in chicken IECs stimulated with CP.

Methods and results: Chicken IECs were stimulated with CP toxin in the absence or presence of antibodies (α-toxin or NetB) for 18 h. Expression of chicken IL-10 and IL-6 was monitored by quantitative real-time polymerase chain reaction. Serum and jejunum samples were collected from CP-infected chickens at 9 days post-infection and expression of IL-10 and IL-6 was analyzed. IL-10 production was detected by antigen capture ELISA in chicken IECs stimulated with CP and in serum samples collected from CP-infected birds. The mRNA levels were consistent with the results of antigen capture ELISA.

Conclusion: CP induced IL-10 production in chicken IECs. Increased IL-10 production was detected in CP-stimulated chicken IECs and in serum collected from CP-infected birds, using antigen capture ELISA. Antigen capture ELISA could be a useful tool to monitor IL-10 production in chicken disease.

Keywords: ELISA; Inflammation; Interleukin-10; Intestinal epithelial cells; Necrotic enteritis.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology
  • Blotting, Western / veterinary
  • Chickens / immunology*
  • Chickens / metabolism
  • Chickens / microbiology
  • Clostridium perfringens / immunology*
  • Enterocolitis, Pseudomembranous / immunology
  • Enterocolitis, Pseudomembranous / microbiology
  • Enterocolitis, Pseudomembranous / pathology
  • Enterocolitis, Pseudomembranous / veterinary*
  • Enzyme-Linked Immunosorbent Assay / methods
  • Enzyme-Linked Immunosorbent Assay / veterinary*
  • Interleukin-10 / metabolism*
  • Intestinal Mucosa / immunology
  • Intestinal Mucosa / metabolism*
  • Intestinal Mucosa / microbiology
  • Necrosis
  • Poultry Diseases / immunology*
  • Poultry Diseases / metabolism
  • Poultry Diseases / pathology
  • Reverse Transcriptase Polymerase Chain Reaction / veterinary

Substances

  • Antibodies, Monoclonal
  • Interleukin-10