Lee Youngsub, Kim Woo H, Lee Sung-Jin, Lillehoj Hyun S
Animal Bioscience and Biotechnology Laboratory, Beltsville Agricultural Research Center, Agricultural Research Service, United States Department of Agriculture, Beltsville, MD, USA.
College of animal Life Sciences, Kangwon National University, Chuncheon 200-701, South Korea.
Vet Immunol Immunopathol. 2018 Oct;204:52-58. doi: 10.1016/j.vetimm.2018.10.001. Epub 2018 Oct 11.
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.
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.
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.
使用针对鸡白细胞介素-10(IL-10)的小鼠单克隆抗体开发的抗原捕获酶联免疫吸附测定(ELISA),评估产气荚膜梭菌(CP)刺激的肠上皮细胞(IECs)和CP感染鸡中IL-10的产生水平。此外,研究在CP刺激的鸡IECs中,哪些CP毒素诱导了IL-10的产生。
在存在或不存在抗体(α毒素或NetB)的情况下,用CP毒素刺激鸡IECs 18小时。通过定量实时聚合酶链反应监测鸡IL-10和IL-6的表达。在感染后9天从CP感染的鸡中收集血清和空肠样本,并分析IL-10和IL-6的表达。通过抗原捕获ELISA在CP刺激的鸡IECs和从CP感染鸟类收集的血清样本中检测到IL-10的产生。mRNA水平与抗原捕获ELISA的结果一致。
CP诱导鸡IECs产生IL-10。使用抗原捕获ELISA在CP刺激的鸡IECs和从CP感染鸟类收集的血清中检测到IL-10产生增加。抗原捕获ELISA可能是监测鸡病中IL-10产生的有用工具。