The Immunoproteome of Francisella Tularensis

导论.土拉热弗朗西丝菌是一种革兰氏阴性兼性胞内病原体， 是一种潜在的生物战剂。令人惊讶的是，人们对整个蛋白质的了解很少。 这种细菌的生理学，只有少数潜在的疫苗候选人被确定在 分子水平。最近的努力是对减毒疫苗株的基因组进行测序 F.土拉热亚种holarctica LVS和强毒F.土拉热亚种土拉菌菌株Schu 4.的 这两种生物体的完整基因组序列使得高通量基因组测序的应用成为可能。 以及用于鉴定疫苗抗原的蛋白质组学技术。具体来说，蛋白质组学技术 包括所开发的阵列技术，其被实施以鉴定其它细胞内抗原的优势抗原， 病原体将被施加到F.土拉热。这些抗原筛选将提供一组确定的潜在 将与佐剂制剂一起评价的候选疫苗， 所引发的免疫应答的功效和性质。该项目的总体目标是开发三个 四种疫苗制剂，适用于GMP下的规模化生产并过渡到临床 问题研究 项目互动：该项目将与其他项目和核心设施紧密结合 在这个提案中描述。这个项目的私家侦探是贝丽尔博士彼得森博士和博西奥将担任共同- Pis和Feigner博士将担任加州大学欧文分校分包合同的PI。我们将 与基因组学/蛋白质组学核心紧密合作，将生物信息学应用于Schu 4 ORF的鉴定， 用质谱法对蛋白质进行分子鉴定。我们也将与动物密切合作。 核心是建立F.土拉热感染，并将转移协议， 疫苗测试在这个核心。随着成功的候选疫苗和制剂的确定， 转移至PDM Core，在GMP条件下进行生产。实施的基本方法 该项目将适用于开发针对其他特定病原体的疫苗。特别是在 将与针对Coxiella spp疫苗开发的团体密切互动（项目 （第二.E.2、第二.E.3和第二.E.4段）以及阐明对伯克霍尔德氏菌属的免疫应答。（项目M.A.4.）。 还将与Kedl博士和Blair博士（项目II.C.6）就脂质体的使用进行密切互动， 用于递送疫苗抗原的载体

Introduction. Francisella tularensis is a gram-negative facultative intracellular pathogen that causes lethal disease in humans, and is a potential biowarfare agent. Surprisingly little is known about the overall protein physiology of this bacterium, and only a handful of potential vaccine candidates are identified at the molecular level. Recent efforts were undertaken to sequence the genomes of the attenuated vaccine strain F. tularensis subsp. holarctica LVS and the virulent F. tularensis subsp. tularensis strain Schu4. The completed genome sequences of these two organisms enable the application of high throughput genomic and proteomic techniques for the identification of vaccine antigens. Specifically, proteomic techniques including array technologies developed implemented to identify dominant antigens of other intracellular pathogens will be applied to F. tularensis. These antigen screens will provide a defined set of potential vaccine candidates that will be evaluated in conjunction with adjuvant formulations to assess protective efficacy and the nature of the immune response elicited. The overall goal of this project is to develop three to four vaccine formulations that are suitable for scale-up production under GMP and transition to clinical studies. Project interactions: This project will be closely integrated with other projects and with Core Facilities described in this proposal. The Pis of this project is Dr. Belisle. Dr. Petersen and Bosio will serve as Co- Pis, and Dr. Feigner will serve as the PI of a sub-contract to the University of California, Irvine. We will work closely with the Genomics/Proteomics Core to apply bioinformatics to the identification of Schu4 ORFs and the molecular identification of proteins by mass spectrometry. We will also work closely with the Animal Core in the implementation of the murine model for F. tularensis infection and will transfer protocols for vaccine testing to this core. As successful vaccine candidates and formulations are identified they will be transferred to the PDM Core for production under GMP conditions. The basic approaches implemented in this project will be applicable to the development of vaccines against other select agents. In particular there will be close interaction with groups that are targeting the development of vaccines to Coxiella spp (Projects II.E.2, II.E.3, and II.E.4.) and the elucidation of the immune response to Burkholderia spp. (Project M.A.4.). There will also be close interaction with Drs. Kedl and Blair (Project II.C.6) on the use of liposomes as a vehicle for the delivery of vaccine antigens