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Oral application of freeze-dried yeast particles expressing the PCV2b Cap protein on their surface induce protection to subsequent PCV2b challenge in vivo

机译:口服应用在其表面表达PCV2b Cap蛋白的冻干酵母颗粒可诱导对体内后续PCV2b攻击的保护

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摘要

Porcine circovirus type 2 (PCV2) is now endemic in every major pig producing country, causing PCV-associated disease (PCVAD), linked with large scale economic losses. Current vaccination strategies are based on the capsid protein of the virus and are reasonably successful in preventing PCVAD but fail to induce sterile immunity. Additionally, vaccinating whole herds is expensive and time consuming. In the present study a “proof of concept” vaccine trial was employed to test the effectiveness of powdered freeze-dried recombinant Saccharomyces cerevisiae yeast stably expressing the capsid protein of PCV2b on its surface as an orally applied vaccine. PCV2-free pigs were given 3 doses of vaccine or left un-vaccinated before challenge with a defined PCV2b strain. Rectal temperatures were measured and serum and faeces samples were collected weekly. At the end of the study, pigs were euthanized, tissue samples taken and tested for PCV2b load by qPCR and immunohistochemistry. The peak of viraemia in sera and faeces of unvaccinated pigs was higher than that of vaccinated pigs. Additionally more sIgA was found in faeces of vaccinated pigs than unvaccinated. Vaccination was associated with lower serum concentrations of TNFα and IL-1β but higher concentrations of IFNα and IFNγ in comparison to the unvaccinated animals. At the end of the trial, a higher viral load was found in several lymphatic tissues and the ileum of unvaccinated pigs in comparison to vaccinated pigs. The difference between groups was especially apparent in the ileum. The results presented here demonstrate a possible use for recombinant S. cerevisiae expressing viral proteins as an oral vaccine against PCV2. A powdered freeze-dried recombinant S. cerevisiae used as an oral vaccine could be mixed with feed and may offer a cheap and less labour intensive alternative to inoculation with the additional advantage that no cooling chain would be required for vaccine transport and storage.
机译:猪圆环病毒2型(PCV2)现在在每个主要的猪生产国都很流行,导致PCV相关疾病(PCVAD),并造成大规模的经济损失。当前的疫苗接种策略是基于病毒的衣壳蛋白,在预防PCVAD方面相当成功,但不能诱导无菌免疫。此外,给整个牛群接种疫苗既昂贵又费时。在本研究中,采用“概念验证”疫苗试验来测试粉状冻干重组酿酒酵母酵母在口服表面稳定表达PCV2b衣壳蛋白的有效性。在用确定的PCV2b毒株攻击之前,给无PCV2的猪接种3剂疫苗或不接种疫苗。测量直肠温度,每周收集血清和粪便样本。在研究结束时,对猪实施安乐死,采集组织样品,并通过qPCR和免疫组织化学测试PCV2b负荷。未接种的猪的血清和粪便中的病毒血症峰值高于未接种的猪。此外,在接种疫苗的猪粪中发现的sIgA高于未接种疫苗的猪。与未接种疫苗的动物相比,接种疫苗可降低血清TNFα和IL-1β的浓度,但可提高IFNα和IFNγ的浓度。在试验结束时,与未接种疫苗的猪相比,未接种疫苗的猪的一些淋巴组织和回肠发现病毒载量更高。两组之间的差异在回肠中尤为明显。此处显示的结果证明,表达重组蛋白的酿酒酵母可能作为抗PCV2的口服疫苗使用。可以将用作口服疫苗的粉状冻干重组酿酒酵母与饲料混合,并且可以提供一种廉价且劳动强度小的替代疫苗,并具有疫苗运输和储存不需要冷却链的额外优势。

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