Development of a Mucosal Vaccine Against Francisella tularensis

土拉弗朗西斯菌粘膜疫苗的研制

基本信息

批准号：
7209733
负责人：
Suzanne M. Michalek
金额：
$ 53.79万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-09-15 至 2009-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7209733
关键词：
Adjuvant Aerosols Animal Model Antibodies Antigen-Presenting Cells Antigens Attenuated Attenuated Live Virus Vaccine Attenuated Vaccines Biological Assay Biological Warfare Bite Categories Cell Proliferation Cells Chaperonin 60 Culicidae Deer Tick Development Disease Effectiveness Enzyme-Linked Immunosorbent Assay Experimental Animal Model Flow Cytometry Food Francisella tularensis GPI-0100 Goals Heat shock proteins Human Immune response Immune system Immunity Immunization In Vitro Infection Infectious Agent Knockout Mice Lipopolysaccharides Measures Mediating Membrane Metabolic Pathway Molecular Chaperones Mus Mutation Nature Penetration Production Purines Ricin Role Route Safety Saponin Saponins Serum Signal Pathway Signal Transduction Soapbush Surface System T-Lymphocyte Testing Tissues Toll-like receptors Tularemia Vaccines Virulence Water analog attenuation chaperonin cytokine fly immunogenicity in vivo microorganism microorganism antigen mucosal vaccine mutant pathogen purine respiratory response shikimate transmission process vaccine development

项目摘要

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）：大多数传染性病原体通过我们的粘膜表面引起疾病，这也适用于生物战制剂。因此，在开发针对传染性/生物战剂的疫苗时，重要的是诱导作用于粘膜表面以及全身区室的反应。土拉弗朗西斯菌是一种革兰氏阴性病原体，也是兔热病的病因。这种微生物是“A 类病原体和生物战剂”。该项目的总体目标是开发一种安全的粘膜疫苗，可有效诱导针对土拉杆菌感染的保护性反应。具体来说，我们将 1) 确定土拉弗拉菌热休克蛋白的免疫原性以及皂苷类似物 GPI-0100 和蓖麻毒素 B 在小鼠全身或鼻内免疫后调节宿主免疫反应中的作用。将收集血清和分泌物并通过 ELISA 测定抗原特异性抗体活性的性质和水平。将培养细胞并评估抗原特异性 T 细胞增殖反应和细胞因子产生（通过 ELISA）。将在用土拉弗拉菌进行全身或粘膜攻击后评估保护反应的有效性。 2) 确定土拉弗拉菌及其脂多糖和佐剂调节宿主反应的细胞机制。 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进行评估。来自正常和 TLR 缺陷小鼠的抗原呈递细胞将在体外受到刺激，并通过流式细胞术评估 MHC 和 B7 表达的变化，并通过 ELISA 评估细胞因子的产生。还将确定参与细胞激活的细胞信号传导途径。 TLR 和 B7 敲除小鼠将用于确定 TLR 和 B7 在土拉弗拉菌及其 LPS 反应中的作用。将如上所述评估体液和细胞反应。土拉弗朗西斯菌的清除率将通过微生物分析来测量。 3) 衍生并表征基因定义的土拉弗拉菌 LVS 减毒株，其莽草酸和/或嘌呤代谢途径发生突变，用作活疫苗。将在小鼠中衍生并测试突变体的安全性、通过微生物分析在宿主组织中的持久性、通过诱导细胞（细胞增殖和细胞因子产生）和体液（通过 ELISA 检测抗体活性的性质和水平）反应的免疫原性，以及诱导保护性的有效性。免疫。这些研究将明确先天性和适应性免疫系统在诱导对土拉弗朗西斯菌的保护性反应中的作用，并将确定一种安全有效的疫苗，以抵抗土拉弗朗西斯菌的粘膜或全身攻击。