A novel cholera subunit vaccine based on 4 colonization factors

描述（由申请人提供）：基于目前对霍乱弧菌（Vc）定植、发病机制和人类对霍乱的免疫反应的了解，提出了一种开发霍乱亚单位疫苗的新方法。目前，在世界上许多霍乱问题严重的地区，口服灭活的全细胞（W-C） Vc疫苗被使用。它所诱导的保护程度取决于接种者的年龄和以前接触过的Vc。灭活的W-C霍乱疫苗不表达感染期间表达的几种Vc保护性抗原的免疫原性水平。为了避免这种情况，我们提出了一种策略，部分基于高度成功的b型流感嗜血杆菌和百日咳嗜血杆菌疫苗，利用与载体蛋白结合的碳水化合物表位，或多种毒力因子和类毒素来实现长期免疫。在动物模型中，针对Vc、LPS和3种定植因子的个体抗体（Abs）具有保护作用。这些Vc毒力因子将用于配置霍乱亚单位疫苗，以诱导对Vc发病机制关键的第一步的集中免疫反应。我们假设一种亚单位霍乱疫苗，由4个保护性Vc定殖/粘附因子组成，一次剂量即可诱导保护性免疫。解毒（det）稻叶脂多糖将结合三种毒力因子之一：毒素共调节菌毛A (TcpA)， TcpF或几丁质结合蛋白A （CBP-A）。载体特异性T细胞的帮助将增强抗lps反应，并有助于诱导保护性的载体特异性抗体。韦德和格兰让他们在霍乱研究方面有丰富的经验。W. Wade博士接受过免疫发病学方面的培训。他率先开发了与载体蛋白结合的合成Vc LPS表位的疫苗接种方案。他是抗vc LPS反应方面的专家。Grandjean博士是一位成功的碳水化合物化学家，他已经产生了Vc LPS：蛋白质偶联物。公共卫生相关性：霍乱仍然是一种每年导致数百万人患病和数千人死亡的疾病。我们建议在鉴定已知霍乱弧菌毒力蛋白TcpA保护表位的基础上，研制一种新的亚单位霍乱疫苗。我们实验室以前的工作表明，LPS和TcpA可以构成霍乱疫苗的基础。我们将继续开发一种霍乱亚单位疫苗，该疫苗基于与引起当前大流行的霍乱弧菌菌株定植所需的3种蛋白质成分结合的解毒lps。

DESCRIPTION (provided by applicant): A novel approach is proposed to develop a cholera subunit vaccine based on the current understanding of Vibrio cholerae (Vc) colonization, pathogenesis, and the human immune response to cholera. Currently, a killed, whole-cell (W-C) Vc vaccine delivered orally is used in much of the world where cholera is problematic. The degree of protection it induces depends on the age and the previous exposure of the vaccinees to Vc. The killed W-C cholera vaccines do not express immunogenic levels of several Vc protective antigens that are expressed during infection. To circumvent this, we propose a strategy based in part on the highly successful H. influenzae type b and B. pertussis vaccines that utilize either carbohydrate epitopes bound to carrier proteins, or multiple virulence factors and a toxoid to achieve long-lasting immunity. Individual antibodies (Abs) to Vc LPS and 3 colonization factors are protective in animal models. These Vc virulence factors will be used to configure a cholera subunit vaccine to induce a concentrated immune response to the critical first step(s) in Vc pathogenesis. We hypothesize that a subunit cholera vaccine, composed of 4 protective Vc colonization/adhesion factors will induce protective immunity with 1 dose. Detoxified (det) Inaba LPS will be conjugated to one of three virulence factors: Toxin Co-Regulated pilus A (TcpA), TcpF or chitin-binding protein A (CBP-A). Carrier-specific T cell help will enhance the anti-LPS response and help for induction of protective carrier-specific Abs. The studies bring together Drs. Wade and Grandjean who have extensive experience in cholera research. Dr. W. Wade is trained in immunopathogenesis. He pioneered the development of the vaccination protocols for synthetic Vc LPS epitopes conjugated to carrier proteins. He is an expert in the anti-Vc LPS response. Dr. Grandjean is an accomplished carbohydrate chemist who has generated Vc LPS:protein conjugates. Public Health Relevance: Cholera is still a disease that sickens millions and kills thousands yearly. We propose to formulate a new subunit cholera vaccine based on identification of protective epitopes in a known virulence proteins of V. cholerae: TcpA. Previous work from our lab has shown that LPS and TcpA can form the basis for a cholera vaccine. We will pursue the development of a cholera subunit vaccine based on detoxified-LPS bound to 3 protein component required for colonization of the strain of V. cholera that is causing the current pandemic.