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

Authors: LEE, YOUNGSUB - US Department Of Agriculture (USDA); KIM, WOOHYUN - US Department Of Agriculture (USDA); LEE, SUNG-JIN - Kangwon National University; Lillehoj, Hyun

Submitted to: Veterinary Immunology and Immunopathology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 8/17/2018
Publication Date: 10/11/2018
Citation: Lee, Y., Kim, W., Lee, S., Lillehoj, H.S. 2018. Detection of chicken interleukin-10 production in epithelial cells and necrotic enteritis induced by Clostridium perfringens using capture-ELISA. Veterinary Immunology and Immunopathology. https://doi.org/10.1016/j.vetimm.2018.10.001.
DOI: https://doi.org/10.1016/j.vetimm.2018.10.001

Interpretive Summary: Necrotic enteritis (NE) is a widespread disease caused by abnormal increase of Clostridium perfringens (CP) colonization in the chicken intestinal tract and costs the US poultry industry > $ 6 billion annually. Understanding the mechanism of immunopathogenesis of NE will lead to better strategies to control this infection. The major pathogen, CP, is known to produce more than 20 toxins and NE is mainly caused by CP strain A. This paper reports that CP A strain is pathogenic and induces proinflammatory cytokines such as IL-6 and IL-8 which cause inflammation. To better understand pathogenesis of NE, ARS scientists developed an in vitro model and tested production of IL-10 by CP. The results showed that IL-6 and IL-10 were expressed when chicken cells were stimulated with a CP toxin by quantitative real-time polymerase chain reaction. Furthermore, novel ELISA was developed to detect IL-10 in serum from NE-infected chickens. This assay system will be valuable to further study the role of NE-associated pathogenic factors and assess cytokine production in biological samples.

Technical Abstract: Aims: To assess the production level of interleukin (IL)-10 in Clostridium perfringens (CP)-treated 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 toxin induced IL-10 in CP-treated chicken IECs. Methods and results: Chicken IECs were stimulated with CP toxin in the absence or presence of antibodies (a-toxin or NetB) for 18 h. Expression of chicken IL-6 and IL-10 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-6 and IL-10 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. IL-10 production was detected in CP-treated chicken IECs and in serum collected from CP-infected birds, using antigen capture ELISA. Antigen capture ELISA could be useful as a new tool to monitor IL-10 production in chicken disease.