Staphylococcal superantigen and anthrax inhibitors

葡萄球菌超抗原和炭疽抑制剂

• 批准号: 8233349
• 负责人: Patrick M Schlievert
• 金额: $ 76.1万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8233349
• 关键词: 3-Dimensional; Affinity; Anthrax disease; Antibiotic Resistance; Area; Bacillus anthracis; Basic Science; Binding; Binding Sites; Categories; Cell Separation; Collaborations; Data; Disease; Dose; Engineering; Enterotoxins; Exotoxins; Foundations; Funding; Genes; Goals; HLA-DR Antigens; Half-Life; Homologous Gene; Human; Immune; In Vitro; Influenza; Intervention; Intravenous; Intravenous Immunoglobulins; Libraries; Lung; Mediating; Molecular; Mutagenesis; Mutation; Oryctolagus cuniculus; Oxygen; Participant; Pathogenesis; Pneumonia; Production; Proteins; Publications; Purpura Fulminans; Records; Research Personnel; Role; Staphylococcal Enterotoxin B; Staphylococcus aureus; Structure; Subgroup; Superantigens; Surface; Symptoms; System; T-Cell Receptor; TNF gene; Technology; Testing; Time; Toxic Shock Syndrome Toxin-1; United States; Virulence; Vitamin K 2; Work; Yeasts; anthrax toxin; biodefense; capsule; cytokine; design; extracellular; in vitro activity; in vivo; inhibitor/antagonist; macrophage; methicillin resistant Staphylococcus aureus; pathogen; prevent; programs; protein-histidine kinase; receptor; receptor binding; small molecule; subcutaneous

Long-term goals are to understand the pathogenesis of methicillin-resistant Staphylococcus aureus (MRSA) that secrete superantigens (SAgs) such as enterotoxin (SE) B or C, SE-like Q, and toxic shock syndrome toxin-1 (TSST-1) and Bacillus anthracis. MRSA are emerging pathogens and their SAgs category B select agents. B. anthracis is a category A select agent. We have assembled two projects that are described below. Project 1. High-Affinity Molecules to Inhibit Superantigen Function. When produced in vivo or when purposely administered, SAgs are exceptionally toxic (TSS symptoms at doses of 0.1 ng/human). Most MRSA associated with post-influenza TSS, necrotizing pneumonia, and purpura fulminans produce SEB, SEC, SE-like Q, and TSST-1. Specific Aim 1. To engineer and characterize V(3-T cell receptor (TCR) regions that bind with high affinity to SAgs and neutralize them, most importantly SEB, SEC, SE-like Q, and TSST-1. Specific Aim 2. To engineer and characterize MHC II regions that bind with high affinity to SAgs and neutralize their activities. Specific Aim 3. To test the ability of soluble Vp-TCRs and MHC Ms to neutralize the activities of SAgs in vitro and in vivo in rabbits. Project. 2: Inhibitors of SrrA-SrrB and BrrA-BrrB Regulated Exotoxin Production. MRSA require oxygen for exotoxin production. This observation led to identification of a "Master Switch" two-component system required for oxygen control of exotoxin production in MRSA. This system, designated srrA-srrB, represses exotoxin production and virulence under conditions of low oxygen. Recently, a homolog in B. anthracis was described-BrrA-BrrB. BrrA-BrrB functions to induce anthrax toxin genes and facilitate disease production. The aims will provide a foundation for interfering with diseases by developing small molecule inhibitors. Specific Aim 4. To characterize SrrB and BrrB as histidine kinases with ability to phosphorylate SrrA and BrrA. Specific Aim 5. To evaluate small molecule inhibitors of SrrB/BrrB to prevent production of SAgs, anthrax toxin, and B. anthracis capsule. Aim 6. To determine the 3-D structures of SrrB and BrrB proteins or extracellular/intracellular domains and combined with menaquinone or inhibitors.

长期目标是了解耐甲氧西林金黄色葡萄球菌 (MRSA) 的发病机制 分泌超抗原 (SAgs)，例如肠毒素 (SE) B 或 C、SE 样 Q 和中毒性休克综合征 toxin-1 (TSST-1) 和炭疽杆菌。 MRSA 是新出现的病原体，其 SAgs B 类选择 代理。炭疽杆菌是 A 类选择病原体。我们已经组装了两个项目，如下所述。 项目 1. 抑制超抗原功能的高亲和力分子。当在体内产生或当 如果有意施用，SAgs 具有极高的毒性（剂量为 0.1 ng/人时会出现 TSS 症状）。最多 与流感后 TSS、坏死性肺炎和暴发性紫癜相关的 MRSA 会产生 SEB， SEC、SE-like Q 和 TSST-1。具体目标 1. 设计和表征 V(3-T 细胞受体 (TCR) 区域 以高亲和力与 SAgs 结合并中和它们，最重要的是 SEB、SEC、SE-like Q 和 TSST-1。 具体目标 2. 设计和表征以高亲和力与 SAgs 结合的 MHC II 区域 中和他们的活动。具体目标 3. 测试可溶性 Vp-TCR 和 MHC Ms 中和 SAgs 在兔体内和体外的活性。项目。 2：SrrA-SrrB 和 BrrA-BrrB 抑制剂的调节 外毒素的产生。 MRSA 需要氧气来产生外毒素。这一观察结果确定了 MRSA 中外毒素产生的氧气控制所需的“主开关”双组分系统。这 系统，指定为 srrA-srrB，在低氧条件下抑制外毒素的产生和毒力。 最近，描述了炭疽芽孢杆菌的同源物——BrrA-BrrB。 BrrA-BrrB 具有诱导炭疽毒素的功能 基因并促进疾病的产生。这些目标将为通过以下方式干预疾病奠定基础： 开发小分子抑制剂。具体目标 4. 将 SrrB 和 BrrB 表征为组氨酸激酶 磷酸化 SrrA 和 BrrA 的能力。具体目标 5. 评估 SrrB/BrrB 小分子抑制剂 防止 SAgs、炭疽毒素和炭疽芽孢杆菌胶囊的产生。目标 6. 确定 3-D 结构 SrrB 和 BrrB 蛋白或细胞外/细胞内结构域并与甲基萘醌或抑制剂组合。