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两种活载体疫苗的开发和评估用于肉鸡控制。

Development and Evaluation of Two Live -Vectored Vaccines for Control in Broiler Chickens.

机构信息

Department of Animal Science, The University of Tennessee, Knoxville, Tennessee.

出版信息

Foodborne Pathog Dis. 2019 Jun;16(6):399-410. doi: 10.1089/fpd.2018.2561. Epub 2019 Mar 13.

Abstract

is the leading bacterial cause of human enteritis in developed countries. Human campylobacteriosis is commonly associated with the consumption of undercooked, contaminated chicken, a natural host of . Thus, the control of colonization in poultry at the farm level would reduce the risk of human exposure to this pathogen. Vaccination is an attractive intervention measure to mitigate in poultry. Our recent studies have demonstrated that the outer-membrane proteins CmeC (an essential component of CmeABC multidrug efflux pump) and CfrA (ferric enterobactin receptor) are feasible candidates for immune intervention against . By targeting these two promising vaccine candidates, live attenuated -vectored vaccines were developed and evaluated in this study. Briefly, the and genes were cloned into expression vector pYA3493 and transferred into serovar Typhimurium χ8914, the USDA licensed live attenuated vaccine strain. The oral live vaccines producing CfrA or CmeC (truncated or full length) were successfully constructed by using delicate molecular manipulation despite the challenge due to the potential toxic effect of the cloned gene product in the host. Expression and membrane localization of the target protein in the vaccines were confirmed by immunoblotting. The efficacies of the two live vaccines that produce full-length CfrA or CmeC were evaluated by using broiler chickens. However, oral vaccination of chickens failed to trigger significant systemic and intestinal mucosal immune responses and, consequently, did not confer protection against colonization chickens. The vaccination regimens of the constructed live -vectored vaccine need to be optimized in future studies.

摘要

空肠弯曲菌是发达国家人类肠炎的主要细菌性病因。人类弯曲菌病通常与食用未煮熟、受污染的鸡肉有关,鸡是 的天然宿主。因此,在农场层面控制家禽中的 定植可以降低人类接触这种病原体的风险。疫苗接种是减轻家禽中 定植的一种有吸引力的干预措施。我们最近的研究表明,外膜蛋白 CmeC(CmeABC 多药外排泵的一个必需组成部分)和 CfrA(铁肠杆菌素受体)是针对 的免疫干预的可行候选物。通过针对这两个有前途的疫苗候选物,本研究开发并评估了活减毒疫苗载体疫苗。简而言之, 和 基因被克隆到表达载体 pYA3493 中,并转移到 血清型 Typhimurium χ8914 中,这是美国农业部许可的活减毒疫苗株。尽管由于克隆基因产物在宿主中的潜在毒性作用而面临挑战,但通过精细的分子操作成功构建了产生 CfrA 或 CmeC(截断或全长)的口服活疫苗。通过免疫印迹证实了疫苗中目标蛋白的表达和膜定位。使用肉鸡评估了产生全长 CfrA 或 CmeC 的两种活疫苗的功效。然而,鸡的口服疫苗接种未能引发显著的全身和肠道黏膜免疫反应,因此不能防止 定植鸡。在未来的研究中,需要优化构建的活疫苗载体疫苗的接种方案。

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