Proteomics to elucidate anthrax and its applications to prevent anthrax

阐明炭疽的蛋白质组学及其预防炭疽的应用

基本信息

批准号：
15390139
负责人：
MAKINO Sou-ichi
金额：
$ 9.73万
依托单位：
Obihiro University of Agriculture Veterinary Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15390139/
关键词：
anthrax virukence infectious diseases プロテオームワクチン

项目摘要

The protective antigen (PA)-based cell free vaccine is only licensed vaccine for human use against Bacillus anthracis infection. Although PA shows strong immunogenicity, the capsule or spore-associated somatic antigens may be important as additional vaccine targets for full protection against anthrax. In this study, we determined the protective effect of spore-associated antigens against the infection. We immunized rabbits with formalin-fixed spores of non-toxigenic, unencapsulated B.anthracis that lacked two virulent plasmids, pXO1 and pXO2, and evaluated the protective effects of the immune antibody. The immunostaining and western blot analysis revealed that the anti-B.anthracis (BA) spore IgG specifically bound to the surface of spores or endospores of B.anthracis, but not to the vegetative cells, or ciosely-related other Bacillus spp. such as B.cereus, B.subtilis, and B.thuringiensis. Passively transferred anti-BA spore IgG protected mice from intraperitoneal challenge with lethal dose of fully virulent B.anthracis spores, and increased survival rate in a dose-dependent manner. The CFU values of spleens and livers of infected mice were significantly lower in antibody-treated mice than those of untreated ones. The anti-BA spore IgG also decreased the number of germinated spores in J774.1 macrophages, suggesting that opsonization of spores promoted phagocytosis and subsequent killing by macrophages. These results indicate the usefulness of spore surface antigens as vaccine targets. In combination with major virulent factors such as PA, spore-associated antigens may offer a safer and more effective multicomponent vaccine for B.anthracis infection.

基于保护性抗原（PA）的无细胞疫苗是唯一获得许可的人类用于对抗炭疽杆菌感染的疫苗。尽管 PA 显示出很强的免疫原性，但荚膜或孢子相关的体细胞抗原作为全面预防炭疽的额外疫苗靶标可能很重要。在这项研究中，我们确定了孢子相关抗原对感染的保护作用。我们用福尔马林固定的无产毒、无包膜的炭疽芽孢杆菌孢子对兔子进行免疫接种，该孢子缺乏两种强毒力质粒 pXO1 和 pXO2，并评估了免疫抗体的保护作用。免疫染色和蛋白质印迹分析表明，抗炭疽芽孢杆菌（BA）孢子 IgG 特异性结合至炭疽芽孢杆菌的孢子或内生孢子表面，但不结合至营养细胞或密切相关的其他芽孢杆菌属。例如蜡状芽孢杆菌、枯草芽孢杆菌和苏云金芽孢杆菌。被动转移的抗 BA 孢子 IgG 可保护小鼠免受致死剂量的全毒力炭疽芽孢杆菌孢子的腹膜内攻击，并以剂量依赖性方式提高存活率。感染小鼠的脾脏和肝脏的CFU值在抗体处理的小鼠中显着低于未处理的小鼠。抗 BA 孢子 IgG 还减少了 J774.1 巨噬细胞中萌发孢子的数量，表明孢子的调理作用促进了巨噬细胞的吞噬作用和随后的杀伤作用。这些结果表明孢子表面抗原作为疫苗靶标的有用性。与 PA 等主要毒力因子相结合，孢子相关抗原可能为炭疽杆菌感染提供更安全、更有效的多组分疫苗。