Identification of protective proteins of Francisella using a novel comparative im

使用新型比较免疫分析方法鉴定弗朗西斯菌的保护蛋白

• 批准号: 8790425
• 负责人: Jyotika Sharma
• 金额: $ 21.54万
• 依托单位: UNIVERSITY OF NORTH DAKOTA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-02-01 至 2018-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8790425
• 关键词: Acute; Aerosols; Animals; Antibodies; Antibody Response; Antigens; Aspartate Transaminase; Attenuated; Automobile Driving; Bacteria; Bacterial Antigens; Breathing; Categories; Centers for Disease Control and Prevention (U.S.); Clinical Research; Collection; Development; Disease; Francisella; Francisella tularensis; Genes; Goals; Health; Immune; Immune response; Immunity; Immunotherapy; Infection; Infection prevention; Inflammatory Response; Lead; Left; Licensing; Life; Lung; Mediating; Membrane; Methods; Mus; Mutant Strains Mice; Organism; Outcome; Peptide Elongation Factor Tu; Prevention strategy; Protein Disulfide Isomerase; Proteins; Proteomics; Pulmonary tularemia; Recombinant Fusion Proteins; Recombinants; Resolution; Respiratory Tract Infections; Route; Serum; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Subunit Vaccines; Testing; Tularemia; Vaccinated; Vaccine Research; Vaccines; Virulence; Virulence Factors; Virulent; Wild Type Mouse; Work; arm; attenuation; base; cell type; comparative; disorder prevention; human disease; in vivo; mortality; mutant; novel; novel strategies; novel vaccines; pathogen; prophylactic; protective efficacy; protein profiling; research study; respiratory; response; vaccine candidate

DESCRIPTION (provided by applicant): Francisella tularensis is the causative agent of respiratory tularemia, a debilitating disease of humans. This bacterial pathogen has been listed as Category A Select Agent owing to its extreme virulence and the ease of its dissemination via aerosol route. To date there is no effective immune therapy or vaccine licensed for prevention of this disease. Although clinical and experimental studies have shown that Th-1 type of host immune responses are protective, bacterial antigens driving these responses are not well defined. Identification of such antigens will aide in formulating effective prevention strategies fr this debilitating disease. This encompasses the goal of the proposed studies. For this, we aim to utilize a novel approach of comparing the immunodominant protein profile of Francisella using sera from mice inoculated with a mutant Francisella strain that is attenuated for causing the infection but does not protect the mice from a lethal challenge with the virulent wild-type organisms and sera from mice inoculated with mutants that not only are attenuated but also protect the mice from lethal challenge. Based on our preliminary studies we believe that this unique approach will identify Francisella proteins associated only with the protective response which can then be utilized as vaccine candidates. In this line, we are armed with a collection of attenuated/non-protective and attenuated/protective mutants of Francisella which will be used to inoculate the mice and the sera collected from these mice will be used to probe total proteins of Francisella followed by sequencing and identification of immunodominant proteins reactive only to the sera from mice inoculated with protective mutants (Aim 1). These proteins will then be produced as recombinant fusion proteins and tested for their protective efficacy against pulmonary infection with virulent Francisella strains (Aim 2). We believe that these studies will uncover Francisella proteins capable of generating protective anti-Francisella immunity thus serving as candidates for a subunit vaccine against this pathogen. Additionally, this novel strategy of comparative immunoproteomics may serve as a platform to identify vaccine candidates for other bacterial pathogens as well. The outcome of proposed studies is expected to take the Francisella subunit vaccine research a step further.

描述（由申请方提供）：土拉热弗朗西斯菌是呼吸道土拉菌病（一种使人衰弱的疾病）的病原体。由于这种细菌病原体毒性极强，而且易于通过气溶胶途径传播，因此被列为A类特定制剂。到目前为止，还没有有效的免疫疗法或疫苗被许可用于预防这种疾病。尽管临床和实验研究表明Th-1型宿主免疫应答具有保护性，但驱动这些应答的细菌抗原尚未明确。这些抗原的鉴定将有助于制定有效的预防策略，这种使人衰弱的疾病。这包括拟议研究的目标。为此，我们的目的是利用一种新的方法，比较免疫显性蛋白质谱的弗朗西斯菌使用血清接种的突变弗朗西斯菌菌株，是减毒的引起感染，但不保护小鼠从致命的挑战与毒性野生型生物体和血清接种的突变体，不仅是减毒的，但也保护小鼠从致命的挑战的小鼠。基于我们的初步研究，我们相信这种独特的方法将鉴定出仅与保护性反应相关的弗朗西斯菌蛋白，然后可以将其用作候选疫苗。在这一系列中，我们配备了一系列弗朗西斯氏菌的减毒/非保护性和减毒/保护性突变体，这些突变体将用于接种小鼠，从这些小鼠收集的血清将用于探测弗朗西斯氏菌的总蛋白，然后测序和鉴定仅与接种保护性突变体的小鼠的血清反应的免疫显性蛋白（目的1）。然后将这些蛋白质作为重组融合蛋白生产，并测试它们对强毒弗朗西斯菌属菌株肺部感染的保护功效（Aim 2）。我们相信，这些研究将揭示弗朗西斯菌蛋白能够产生保护性抗弗朗西斯菌免疫，从而作为针对这种病原体的亚单位疫苗的候选者。此外，这种比较免疫蛋白质组学的新策略也可以作为鉴定其他细菌病原体的候选疫苗的平台。拟议研究的结果有望使弗朗西斯菌亚单位疫苗研究更进一步。