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棘球蚴多房绦虫表面糖蛋白的特性及其在犬中的黏膜疫苗潜力。

Characterization of a surface glycoprotein from Echinococcus multilocularis and its mucosal vaccine potential in dogs.

机构信息

Department of Infectious Diseases, Hokkaido Institute of Public Health, Sapporo, Japan.

出版信息

PLoS One. 2013 Jul 23;8(7):e69821. doi: 10.1371/journal.pone.0069821. Print 2013.

Abstract

Alveolar echinococcosis is a refractory disease caused by the metacestode stage of Echinococcus multilocularis. The life cycle of this parasite is maintained primarily between foxes and many species of rodents; thus, dogs are thought to be a minor definitive host except in some endemic areas. However, dogs are highly susceptible to E. multilocularis infection. Because of the close contact between dogs and humans, infection of dogs with this parasite can be an important risk to human health. Therefore, new measures and tools to control and prevent parasite transmission required. Using 2-dimensional electrophoresis followed by western blot (2D-WB) analysis, a large glycoprotein component of protoscoleces was identified based on reactivity to intestinal IgA in dogs experimentally infected with E. multilocularis. This component, designated SRf1, was purified by gel filtration using a Superose 6 column. Glycosylation analysis and immunostaining revealed that SRf1 could be distinguished from Em2, a major mucin-type antigen of E. multilocularis. Dogs (n=6) were immunized intranasally with 500 µg of SRf1 with cholera toxin subunit B by using a spray syringe, and a booster was given orally using an enteric capsule containing 15 mg of the same antigen. As a result, dogs immunized with this antigen showed an 87.6% reduction in worm numbers compared to control dogs (n=5) who received only PBS administration. A weak serum antibody response was observed in SRf1-immunized dogs, but there was no correlation between antibody response and worm number. We demonstrated for the first time that mucosal immunization using SRf1, a glycoprotein component newly isolated from E. multilocularis protoscoleces, induced a protection response to E. multilocularis infection in dogs. Thus, our data indicated that mucosal immunization using surface antigens will be an important tool to facilitate the development of practical vaccines for definitive hosts.

摘要

泡球蚴病是由细粒棘球绦虫的中绦期幼虫引起的难治性疾病。这种寄生虫的生命周期主要在狐狸和许多种啮齿动物之间维持;因此,除了在一些流行地区外,狗被认为是次要的终末宿主。然而,狗非常容易感染细粒棘球蚴。由于狗与人之间的密切接触,狗感染这种寄生虫可能是对人类健康的一个重要威胁。因此,需要采取新的措施和工具来控制和预防寄生虫传播。通过二维电泳结合免疫印迹(2D-WB)分析,根据对实验感染细粒棘球蚴的狗肠 IgA 的反应性,鉴定出原头蚴的一个大型糖蛋白成分,命名为 SRf1。该成分通过 Superose 6 柱凝胶过滤进行纯化。糖基化分析和免疫染色表明,SRf1 可以与 Em2 区分开来,Em2 是细粒棘球蚴的主要粘蛋白型抗原。用喷雾注射器鼻内免疫 6 只狗 500 µg 的 SRf1 与霍乱毒素亚单位 B,并用肠溶胶囊口服给予同一抗原 15mg 进行加强免疫。结果,与仅接受 PBS 给药的对照组狗(n=5)相比,用这种抗原免疫的狗的虫体数量减少了 87.6%。在 SRf1 免疫的狗中观察到较弱的血清抗体反应,但抗体反应与虫体数量之间没有相关性。我们首次证明,使用从细粒棘球蚴原头节中分离出的糖蛋白成分 SRf1 进行黏膜免疫可诱导狗对细粒棘球蚴感染产生保护反应。因此,我们的数据表明,使用表面抗原进行黏膜免疫将是开发针对终末宿主的实用疫苗的重要工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4d03/3720955/94bfe6e15641/pone.0069821.g001.jpg

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