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Identification of secreted proteins as novel antigenic vaccine candidates of Haemophilus parasuis serovar 5

机译：鉴定分泌蛋白作为副猪嗜血杆菌血清型5的新型抗原候选疫苗

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Haemophilus parasuis (H. parasuis) is a swine pathogen responsible for the Glasser's disease, which has received more attention in the past decade due to the increasing economic losses in the pig industry worldwide. As traditional inactive vaccine of H. parasuis has obvious disadvantage, to identify efficient immunoprotective antigens would undoubtedly contribute to the development of novel subunit vaccines. The putative secreted proteins of H. parasuis are potentially essential components of more potent vaccines. In the present study, six secreted proteins (MA, Gcp, Ndk, HsdS, RnfC and HAPS_0017) were selected from the annotated H. parasuis serovar 5 genome as immunogenic protein with bioinformatic and experimental approaches. These proteins were successfully expressed in Escherichia coli and their immunogenicity was assessed in a mouse challenge model. The results showed that subcutaneous injection with the recombinant proteins resulted in the production of antibodies with high levels. Antigen-specific lymphoproliferative responses were detected in the splenocytes of the immunized animals. CD4(+) T-cell populations were higher in the vaccinated animals 3 weeks after the booster immunization than those of the control animals. A significant increase was observed in the cytokine levels of IL-2, IL-4 and IFN-gamma in the culture supernatants of splenocytes. Furthermore, immunized mice conferred different levels of protection against challenge with a lethal dose of highly virulent serovar 5 strain (H46). Our results indicate that these six secreted proteins induced a good Th1 response and protection against H. parasuis infection, could be potential subunit vaccine candidates. (C) 2015 Elsevier Ltd. All rights reserved.
机译：副猪嗜血杆菌（H. parasuis）是造成格拉瑟氏病的猪病原体，由于全球养猪业的经济损失日益增加，近十年来受到越来越多的关注。由于传统的副猪嗜血杆菌无活性疫苗具有明显的缺点，因此鉴定有效的免疫保护性抗原无疑将有助于新型亚单位疫苗的开发。副猪嗜血杆菌的推定分泌蛋白可能是更有效疫苗的潜在重要组成部分。在本研究中，通过生物信息学和实验方法从带注释的副猪嗜血杆菌血清型5基因组中选择了六个分泌蛋白（MA，Gcp，Ndk，HsdS，RnfC和HAPS_0017）作为免疫原性蛋白。这些蛋白质已在大肠杆菌中成功表达，并在小鼠攻击模型中评估了其免疫原性。结果表明，皮下注射重组蛋白导致高水平抗体的产生。在免疫动物的脾细胞中检测到抗原特异性淋巴增生反应。加强免疫后3周，接种动物的CD4（+）T细胞数量高于对照组动物。在脾细胞培养上清液中观察到IL-2，IL-4和IFN-γ的细胞因子水平显着增加。此外，免疫小鼠赋予了致命剂量的高毒力血清型5株（H46）抵抗攻击的不同水平的保护。我们的结果表明，这六个分泌蛋白诱导了良好的Th1反应，并能抵抗副猪嗜血杆菌感染，可能是潜在的亚单位疫苗候选药物。 （C）2015 Elsevier Ltd.保留所有权利。

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