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Evaluation of in ovo vaccination of DNA vaccines for Campylobacter control in broiler chickens

机译：评价DNA疫苗植物疫苗植物植物鸡肉鸡

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Campylobacter is the leading bacterial cause of human enteritis in developed countries. Chicken is a major natural host of Campylobacter. Thus, on-farm control of Campylobacter load in poultry would reduce the risk of human exposure to this pathogen. Vaccination is an attractive intervention measure to mitigate Campylobacter in poultry. Our previous studies have demonstrated that Campylobacter outer membrane proteins CmeC (a component of multidrug efflux pump) and CfrA (ferric enterobactin receptor) are feasible and promising candidates for vaccine development. In this study, by targeting these two attractive vaccine candidates, we explored and evaluated a new vaccination strategy, which combines the in ovo vaccination route and novel DNA vaccine formulation, for Campylobacter control in broilers. We observed that direct cloning of cfrA or cmeC gene into the eukaryotic expression vector pCAGGS did not lead to sufficient level of production of the target proteins in the eukaryotic HEK-293 cell line. However, introduction of the Kozak consensus sequence (ACCATGG) in the cloned bacterial genes greatly enhanced production of inserted gene in eukaryotic cells, creating desired DNA vaccines. Subsequently, the validated DNA vaccines were prepared and used for two independent in ovo vaccination trials to evaluate their immune response and protective efficacy. However, single in ovo injection of specific DNA vaccine at 18th day of embryonation, regardless using neutral lipid-protected vector or not, failed to trigger significant IgG and IgA immune responses and did not confer protection against C. jejuni colonization in the intestine of chickens. In conclusion, this study demonstrates that the Kozak sequence is critically important for construction of the DNA vaccine expressing prokaryotic gene. The optimal regimen for in ovo vaccination of DNA vaccine for Campylobacter control in poultry needs to be determined in future studies. (C) 2019 Elsevier Ltd. All rights reserved.
机译：Campylobacter是发达国家人类肠炎的主要细菌原因。鸡是一个主要的弯曲杆菌的主要自然宿主。因此，家禽弯曲杆菌载荷的农场控制会降低人体暴露于该病原体的风险。疫苗接种是一种吸引人的干预措施，可以减轻家禽的弯曲杆菌。我们以前的研究表明，弯曲杆菌外膜蛋白CMEC（多药渗透泵的组分）和CFRA（肠肠球受体）是疫苗发育的可行和有希望的候选者。在这项研究中，通过靶向这两个吸引力的疫苗候选者，我们探讨并评估了一种新的疫苗接种策略，它结合了卵巢疫苗接种途径和新型DNA疫苗配方，用于肉鸡中的弯曲杆菌控制。我们观察到，CFRA或CMEC基因将CFRA或CMEC基因的直接克隆到真核表达载体PCAGG中没有导致真核安全性HEK-293细胞中靶蛋白的充分产生水平。然而，在克隆的细菌基因中引入Kozak共有序列（Accatgg）大大增强了真核细胞中插入基因的生产，产生了所需的DNA疫苗。随后，制备验证的DNA疫苗并用于两个在OVO疫苗接种试验中的两个独立的，以评估它们的免疫应答和保护效果。然而，在胚胎中的18℃注入特定的DNA疫苗，无论使用中性的脂质保护载体，都未能引发显着的IgG和IgA免疫反应，并且在鸡肠中没有赋予C. Jejuni定植的保护。 。总之，本研究表明，Kozak序列对表达原核基因的DNA疫苗构建至关重要。在未来的研究中，需要确定在家禽中用于弯曲杆菌的DNA疫苗疫苗接种的最佳方案需要在未来的研究中确定。 （c）2019 Elsevier Ltd.保留所有权利。

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