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使用捕获ELISA检测产气荚膜梭菌诱导的鸡肠道上皮细胞中白细胞介素-10的产生及坏死性肠炎

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

作者信息

Lee Youngsub, Kim Woo H, Lee Sung-Jin, Lillehoj Hyun S

机构信息

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.

出版信息

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.

摘要

目的

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。

方法与结果

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。

结论

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。

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