Construction of safe and effective live tuberculosis vaccines

安全有效的结核活疫苗的构建

基本信息

批准号：
8626351
负责人：
Steven A Porcelli
金额：
$ 41.5万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-03-01 至 2016-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8626351
关键词：
Address Adjuvant Animals Antigen Presentation Antigens Apoptosis Apoptotic Attenuated Attenuated Vaccines Bacillus (bacterium)Bacteriophages CD4 Positive T Lymphocytes CD8B1 gene Cells Chemicals Communicable Diseases Cosmids Disease Gene Deletion Gene Mutation Genes Genetic Goals Growth Histocompatibility Antigens Class I Histocompatibility Antigens Class II Human Immune Immune response Immunity Immunologic Deficiency Syndromes Immunology In Vitro Laboratories Lead Life Link MHC Class II Genes Mediating Metabolic Methodology Multi-Drug Resistance Mus Mutagenesis Mutation Mycobacterium smegmatis Mycobacterium tuberculosis Phagocytes Population Post-Translational Protein Processing Prevention Production Protein Subunits Proteins Research Personnel Rodent Specificity System T cell response T memory cell T-Lymphocyte T-Lymphocyte Epitopes T-Lymphocyte Subsets TNF gene Testing Tuberculosis Tuberculosis Vaccines Vaccination Vaccines Virulence Work auxotrophy cytokine design and construction human disease immunogenic immunogenicity improved in vivo interest mortality mouse model mutant mycobacterial novel overexpression pathogen pre-clinical prevent public health relevance resistant strain response screening vaccine candidate vaccine development

项目摘要

DESCRIPTION (provided by applicant): This proposal seeks to understand in greater detail the mechanisms by which Mycobacterium tuberculosis blocks the priming of effective host immunity, with the ultimate goal of using this information to create more effective live vaccine strains. Previous work identified multiple mycobacterial genes involved in blocking apoptosis of infected host cells, which is intimately linked to the ability of the pathogen to prevent presentation of its antigens by MHC class I. Extensive preliminary work has also identified genes in M. tuberculosis that interfere with MHC class II antigen presentation, and genes that block production of key cytokines. A major goal of this proposal is to construct safely attenuated strains of M. tuberculosis in which specific immune evasion genes have been deleted, thus creating more effective vaccines for priming of anti- mycobacterial immunity. The approach involves identifying the most potent anti- apoptotic genes from a group of four candidates that have already been partially characterized, and combining mutations in these with strongly attenuating auxotrophy mutations to eliminate virulence. Precise mutations in the genes of interest will be created using allelic exchange mediated by specialized transducing phages, a methodology that is well established in the laboratory of the PI and his collaborators. The potency of candidate vaccine strains will be further enhanced by incorporating additional mutations in genes that interfere with MHC class II presentation, or with production of IL-12p70 or TNF. Immunological studies of CD4 and CD8 T cell priming by candidate vaccine strains will be carried out in mouse models, allowing identification of the most favorable combinations of specific gene deletions to advance into preclinical vaccination studies in rodents. Studies of the induction of stable T cell memory by candidate vaccine strains will be performed in mice, and the impact of boosting primary responses with mycobacterial proteins will also be initiated. Overall, the proposed studies will significantly advance our understanding of the host-pathogen interaction in tuberculosis, and contribute directly to vaccine development for prevention and control of this major human disease.

描述（由申请人提供）：该提案旨在更详细地了解结核分枝杆菌阻止有效宿主免疫的启动的机制，其最终目标是使用此信息创建更有效的实时疫苗菌株。先前的工作确定了涉及阻断受感染宿主细胞凋亡的多个分枝杆菌基因，这与病原体防止MHC类I级抗原的能力密切相关。广泛的初步工作也确定了结核分枝杆菌中的基因，这些基因与MHC II Antigen Claben呈现的基因构成了Cyper ander ander shiptation和Key key Cytok cytok of Key Cytok的生产。该提案的一个主要目的是构建已删除特定免疫逃避基因的结核分枝杆菌菌株的安全菌株，从而创造出更有效的疫苗来启动抗分枝杆菌免疫。该方法涉及从已经被部分表征的四个候选者中鉴定出最有效的抗凋亡基因，并将突变与强烈衰减的辅助营养突变结合在一起以消除毒力。感兴趣基因的精确突变将使用由专门的转导噬菌体介导的等位基因交换来创建，该方法在PI及其合作者实验室中良好确定。通过在干扰MHC II类表现或产生IL-12P70或TNF的基因中，将候选疫苗菌株的效力进一步增强。候选疫苗菌株对CD4和CD8 T细胞启动的免疫学研究将在小鼠模型中进行，从而可以鉴定出特定基因缺失的最有利组合，从而将其促进啮齿动物的临床前疫苗接种研究。将在小鼠中对候选疫苗菌株诱导稳定的T细胞记忆的诱导研究，并且还将启动用分枝杆菌蛋白增强主要反应的影响。总体而言，拟议的研究将显着提高我们对结核病中宿主 - 病原体相互作用的理解，并直接为预防和控制这种主要人类疾病的疫苗开发做出贡献。