Development of a Mucosal Vaccine Against F. tularensis

土拉弗朗西斯粘膜疫苗的研制

• 批准号: 6689492
• 负责人: Suzanne M. Michalek
• 金额: $ 29.96万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-15 至 2008-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6689492
• 关键词: Francisella tularensis; T lymphocyte; antigen presenting cell; bacterial vaccines; biological signal transduction; biotechnology; bioterrorism /chemical warfare; cellular immunity; cooperative study; cytokine; enzyme linked immunosorbent assay; flow cytometry; gene mutation; gene targeting; genetic strain; heat shock proteins; humoral immunity; immunomodulators; laboratory mouse; laboratory rabbit; lipopolysaccharides; lymphocyte proliferation; mucosal immunity; ricin; shikimate; tissue /cell culture; toll like receptor; tularemia; vaccine development

DESCRIPTION (provided by applicant): Most infectious agents cause disease via our mucosal surfaces, which also applies to biological warfare agents. Therefore, in developing a vaccine against infectious/biological warfare agents, it is important to induce responses that would act at mucosal surfaces, as well as in the systemic compartment. Francisella tularensis is a gram-negative pathogen and cause of tularemia. This microorganism is a "category A pathogen and biological warfare agent". The overall goal of this project is to develop a safe mucosal vaccine effective in inducing protective responses against infection by F. tularensis. Specifically, we will 1) Determine the immunogenicity of heat shock proteins of F. tularensis and the effect of the saponin analog GPI-0100 and of ricin B in modulating host immune responses following systemic or intranasal immunization of mice. Serum and secretions will be collected and assayed for the nature and level of antigen-specific antibody activity by ELISA. Cells will be cultured and assessed for antigen-specific T cell proliferative responses and cytokine production (by ELISA). The effectiveness of the response on protection will be assessed following systemic or mucosal challenge with F. tularensis. 2) Determine the cellular mechanism(s) by which F. tularensis and its LPS, and the adjuvants modulate host responses. The role of Toll-like receptors (TLRs) and the B7 costimulatory system in mediating host responses and infection will be assessed in vitro and in vivo. Antigen-presenting cells from normal and TLR deficient mice will be stimulated in vitro and assessed for changes in the expression of MHC and B7 by flow cytometry and for cytokine production by ELISA. The cell signaling pathways involved in cell activation will also be determined. TLR- and B7-knockout mice will be used to determine the role of TLRs and B7 in responses to F. tularensis and its LPS. Humoral and cellular responses will be assessed as indicated above. Clearance of F. tularensis will be measured by microbiologic analysis. 3) Derive and characterize genetically defined attenuated strains of F. tularensis LVS with mutations in the shikimate and/or purine metabolic pathways for use as a live vaccine. Mutants will be derived and tested in mice for their safety, persistence in host tissue by microbiologic analysis, immunogenicity by inducing cellular (cell proliferation and cytokine production) and humoral (nature and level of antibody activity by ELISA) responses, and effectiveness in inducing protective immunity. These studies will define the role of the innate and adaptive immune systems in inducing protective responses to F. tularensis and will define a safe and efficacious vaccine against mucosal or systemic challenge with F. tularensis.

描述（由申请人提供）：大多数传染剂通过我们的粘膜表面引起疾病，这也适用于生物战剂。 因此，在开发针对感染/生物战剂的疫苗时，重要的是诱导将在粘膜表面以及系统隔室中起作用的反应。 Francisella tularensis是一种革兰氏阴性的病原体，是t骨的原因。这种微生物是“病原体和生物战剂类别”。该项目的总体目标是开发一种安全的粘膜疫苗，可有效诱导F. tolarensis感染的保护性反应。具体而言，我们将1）确定F. tularensis的热休克蛋白的免疫原性，以及小鼠的全身性或鼻内免疫免疫后，皂苷类似物GPI-0100和Ricin B对调节宿主免疫反应的影响。将通过ELISA收集和分泌分泌物，以针对抗原特异性抗体活性的性质和水平进行测定。细胞将进行培养并评估抗原特异性T细胞增殖反应和细胞因子产生（通过ELISA）。在针对F. tularensis的全身性或粘膜挑战之后，将评估反应对保护的有效性。 2）确定F. tolarensis及其LPS及其LPS的细胞机制，以及佐剂调节宿主反应。 Toll样受体（TLR）和B7共刺激系统在介导宿主反应和感染中的作用将在体外和体内评估。来自正常和TLR缺乏小鼠的抗原呈递细胞将在体外刺激，并评估通过流式细胞仪和ELISA的细胞因子产生的MHC和B7表达的变化。还将确定参与细胞激活的细胞信号通路。 TLR-和B7敲除小鼠将用于确定TLR和B7在对F. tularensis及其LPS的响应中的作用。如上所述，将评估体液和细胞反应。 F. tularensis的清除将通过微生物学分析来测量。 3）得出并表征了遗传定义的tularensis lvs的遗传定义的衰减菌株，其在寒气和/或嘌呤代谢途径中具有突变，以用作活疫苗。突变体将通过微生物学分析，通过诱导细胞（细胞增殖和细胞因子产生）以及体液（ELISA的性质和抗体活性水平），通过微生物学分析，通过微生物组织在宿主组织中的持久性来得出突变体，以及诱导保护性免疫的有效性。这些研究将定义先天和适应性免疫系统在诱导对f菌的保护性反应中的作用，并将定义一种安全有效的疫苗，以防止粘膜或针对F. tolarensis的粘膜或系统性挑战。