CHOLERA-VECTORED DELIVERY SYSTEM FOR PERTUSSIS ANTIGENS

霍乱载体百日咳抗原递送系统

基本信息

批准号：
2671410
负责人：
JOAN R BUTTERTON
金额：
$ 8.99万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
1996
资助国家：
美国
起止时间：
1996-05-01 至 1999-04-30
项目状态：
已结题

项目摘要

The applicant proposes to develop the use of Vibrio cholerae, the causative agent of cholera, as a live oral attenuated vaccine vector that can deliver heterologous antigens to stimulate a common mucosal immune response. Bordetella pertussis, the causative agent of whooping cough, is an important pathogen of humans which, like V. cholerae, acts at the mucosal surface. The investigators hypothesize that oral immunization with V. cholerae vector strains expressing protective antigens of B. pertussis may lead to protective immunity against B. pertussis infection in the respiratory tract. Examination of immune responses following infection with the vector strains may aid in elucidating the mechanisms of protective immunity to B. pertussis and may further the understanding of protective immunity at mucosal surfaces. The filamentous hemagglutinin (FHA) of B. pertussis will he used as a model heterologous antigen. FHA is a 220-kDa filamentous protein that is proposed to be one of the major adhesins mediating the interaction of B. pertussis and human cilia. The cloned gene encoding FHA (FhaB) will be placed under the control of an in-vivo expressed V. cholerae promoter. The promoter for the V. cholerae heat-shock gene htpG will be used; this promoter has led to expression of other heterologous antigens from V. cholerae in prior studies. The expression of FHA from htpGp will be compared with that from a constitutively-expressed promoter. The fhaB constructs will be placed onto the chromosome of V. cholerae by in-vivo marker exchange, and the constructs will be tested for expression of the heterologous antigen. A germ-free adult mouse model of colonization by V. cholerae will be developed that can be used to assess the immune response to FHA expressed by the V. cholerae strains, and the induction of protective immunity will be examined using an inhalation challenge model of B. pertussis infection. The Principal Investigator is applying for three additional years of research training in order to further develop the use of V cholerae as a vaccine vector for delivering heterologous antigens. The proposed project will require learning more sophisticated molecular biologic techniques, mastering methods for performing immunologic assays, and working with a variety of new animal models for evaluating colonization, immune response, and protection by the constructed vaccines. The additional research time would allow a smooth transition from post-doctoral training to an independent research career in academic medicine in the specialty of infectious disease. The sponsors are experienced in the necessary techniques and have the resources needed for successful completion of the proposed project.

申请人提议开发霍乱弧菌的用途，霍乱的病原体，作为活口服减毒疫苗载体可以传递异源抗原来刺激共同的粘膜免疫回复。百日咳博德特氏菌（Bordetella pertussis）是百日咳的病原体，人类的一种重要病原体，与霍乱弧菌一样，作用于粘膜表面。研究人员假设口服免疫表达百日咳博德特氏菌保护性抗原的霍乱弧菌载体菌株可能会导致针对百日咳博德特氏菌感染的保护性免疫力呼吸道。感染后免疫反应的检查与载体菌株的结合可能有助于阐明其机制对百日咳博德特氏菌的保护性免疫力，可能会进一步了解粘膜表面的保护性免疫。百日咳博德特氏菌的丝状血凝素（FHA）将被用作模型异源抗原。 FHA 是一种 220 kDa 的丝状蛋白，被认为是介导 B 相互作用的主要粘附素之一。百日咳和人类纤毛。编码 FHA (FhaB) 的克隆基因将是置于体内表达的霍乱弧菌启动子的控制下。这将使用霍乱弧菌热休克基因 htpG 的启动子；这启动子导致V的其他异源抗原的表达。先前研究中的霍乱。 htpGp 的 FHA 表达为与组成型表达的启动子相比。法哈B 构建体将通过体内方法放置到霍乱弧菌的染色体上标记交换，并且将测试构建体的表达异源抗原。 V. 定植的无菌成年小鼠模型。将开发出可用于评估免疫反应的霍乱弧菌霍乱弧菌菌株表达的 FHA，以及诱导将使用吸入激发模型检查保护性免疫力百日咳杆菌感染。首席研究员正在申请额外三年的研究研究培训，以进一步开发霍乱弧菌作为用于传递异源抗原的疫苗载体。拟议项目需要学习更复杂的分子生物学技术，掌握进行免疫测定的方法，并与各种新的动物模型，用于评估定植、免疫反应、和构建疫苗的保护。额外的研究时间实现从博士后培训到博士后培训的顺利过渡学术医学专业的独立研究生涯传染病。赞助商在必要的方面拥有丰富的经验技术并拥有成功完成任务所需的资源拟议的项目。