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（由申请人提供）：弗朗西斯氏菌是呼吸道疾病的病因，呼吸道疾病是一种令人衰弱的人类疾病。由于其极端的毒力以及通过气溶胶途径散布的，该细菌病原体已被列为一种选择剂。迄今为止，还没有有效的免疫治疗或用于预防这种疾病的疫苗。尽管临床和实验研究表明，宿主免疫反应的TH-1类型是保护性的，但驱动这些反应的细菌抗原尚未很好地定义。这种抗原的鉴定将有助于制定这种使人衰弱的疾病的有效预防策略。这涵盖了拟议的研究的目标。为此，我们旨在利用一种新的方法来比较弗朗西斯菌的免疫蛋白质蛋白质的特征，使用接种的小鼠接种了一种与突变的francisella菌株接种的小鼠，该菌株因造成感染而受到衰减，但并不能保护小鼠免受与疾病的野生型生物的无致命挑战，而不仅仅是从事突变型的鼠标，但也不仅保护了菌群。基于我们的初步研究，我们认为这种独特的方法将识别仅与保护性反应相关的francisella蛋白，然后可以将其用作疫苗候选物。 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).然后，这些蛋白质将作为重组融合蛋白产生，并测试其针对用毒francisella菌株的肺部感染的保护作用（AIM 2）。我们认为，这些研究将发现能够产生保护性抗氟西氏菌免疫的弗朗西斯氏菌蛋白，从而作为针对这种病原体的亚基疫苗的候选。此外，这种比较免疫蛋白质组学的新型策略也可以作为识别其他细菌病原体疫苗的平台。预计拟议的研究的结果将进一步将弗朗西斯菌亚基疫苗研究进一步。