Staphylococcal Superantigen Interactions With Vaginal Epithelium

葡萄球菌超抗原与阴道上皮的相互作用

• 批准号: 7728629
• 负责人: Marnie L Peterson
• 金额: $ 29.52万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-13 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7728629
• 关键词: Affect; Amino Acids; Area; Asthma; Atopic Dermatitis; Bacteria; Binding; Biological Models; Blood Circulation; Cells; Collaborations; Cytolysins; Cytotoxin; Data; Dermatitis; Development; Disease; Dose; Environment; Epithelial; Epithelial Cells; Epithelial Receptor Cell; Epithelium; Event; Exotoxins; Exposure to; Family suidae; Fever; Goals; Human; Immune system; Infection; Inflammation; Inflammatory; Intestines; Iowa; Knock-out; Knowledge; Lead; Life; Mediating; Microbe; Minnesota; Modeling; Mucous Membrane; Organism; Pathogenesis; Pathology; Penetration; Permeability; Pneumonia; Principal Investigator; Production; Property; Proteins; Receptor Cell; Reporting; Research; Research Personnel; Resources; Rhinitis; Role; Secondary to; Shock; Signal Pathway; Signal Transduction; Signaling Molecule; Site; Skin; Soft Tissue Infections; Staphylococcus aureus; Streptococcal Infections; Structure; Submucosa; Superantigens; Surface; Syndrome; T-Cell Activation; T-Lymphocyte; Testing; Therapeutic; Thick; Tissues; Toxic Shock Syndrome; Toxic Shock Syndrome Toxin-1; Toxic effect; Toxin; United States; Vagina; Work; base; cytokine; cytotoxic; experience; human disease; improved; innovation; interest; macrophage; microbial; mutant; novel; novel strategies; novel therapeutics; perforin; prevent; programs; public health relevance; receptor; receptor binding; response; therapeutic development

DESCRIPTION (provided by applicant): The long-term goals of this research are to understand how Staphylococcus aureus and its superantigens (SAgs), cytotoxins, and other exoproteins interact with mucosal surfaces to facilitate SAg penetration to cause serious human illnesses. We propose to use the vaginal epithelium as the model multi-layered mucosal surface, and hypothesize the data obtained in planned studies will also be applicable to other mucosal surfaces. Prior results using an ex vivo porcine vaginal permeability model indicate that the SAg, TSST-1, traverses mucosal surfaces in small amounts, but localizes in higher amounts within the inner epithelium, possibly serving as a toxin reservoir. In the presence of staphylococcal 1-toxin, TSST-1 penetrated the full thickness of the vaginal mucosa in significantly higher amounts, with considerable TSST- 1 also remaining within the epithelium. We hypothesize that cytotoxic and inflammatory properties of staphylococcal cytotoxins and other exoproteins enhance penetration of TSST-1 to inner epithelial cells where TSST-1 binds to an undescribed epithelial cell receptor, triggering pro-inflammatory cytokine production. Base on these hypotheses we propose two specific aims: 1) To determine the contribution of S. aureus cytotoxins (exoproteins) in enhancing the penetration of TSST-1 across epithelial surfaces by causing cytolytic and/or inflammatory effects on epithelial cells, and 2) To determine TSST-1 receptor binding and the mechanism of TSST-1 stimulation of epithelial cells, leading to intracellular activation of signaling pathways with subsequent cytokine production. Our preliminary data suggest that the most likely candidate exoproteins to facilitate penetration of TSST-1 across mucosal barriers are cytotoxins, including 3-toxin, which we propose to study in depth through use of purified 3-toxin components and allelic replacement knockout strains. In the course of these studies, we will also study mucosal penetration and activities of a deletion [missing amino acids 1 to 72] mutant TSST-1 protein (13,000 MW vs. 22,000 MW wild-type TSST-1) that maintains superantigenicity and is associated with a rapidly progressive, 100% fatal, extreme pyrexia syndrome in humans. We believe the planned studies will clarify the mechanisms by which TSST-1 penetrates mucosal barriers to cause large numbers of human diseases. These studies may suggest novel therapeutic strategies to manage serious S. aureus illnesses. PUBLIC HEALTH RELEVANCE: The bacterium Staphylococcus aureus often initiates serious human diseases from the organism's colonization of mucous membranes, followed by secretion of potent exotoxins referred to as superantigens. Illnesses include toxic shock syndrome that may have both menstrual and non- menstrual forms, and that may affect thousands of humans each year in the United States. This application studies the mechanism by which superantigens penetrate mucosal surfaces, allowing S. aureus to cause life threatening toxic shock syndrome, and possibly suggesting novel strategies to prevent and manage such illnesses.

描述（由申请人提供）：这项研究的长期目标是了解金黄色葡萄球菌及其超抗原（SAGS），细胞毒素和其他脱甲蛋白如何与粘膜表面相互作用，以促进下垂渗透以引起严重的人类疾病。我们建议将阴道上皮用作模型多层粘膜表面，并假设计划研究中获得的数据也将适用于其他粘膜表面。使用离体猪的阴道通透性模型的先前结果表明，SAG（TSST-1）以少量遍历粘膜表面，但内部上皮内的含量较高，可能是毒素储层。在存在葡萄球菌1毒素的情况下，TSST-1以明显更高的量穿透了阴道粘膜的全部厚度，上皮内也存在相当大的TSST-1。我们假设葡萄球菌细胞毒素和其他销食蛋白的细胞毒性和炎症特性增强了TSST-1的渗透到内皮细胞中，在这些细胞中TSST-1与未描述的上皮细胞受体结合，从而触发持炎性细胞因子的产生。基于这些假设，我们提出了两个具体的目的：1）确定金黄色葡萄球菌细胞毒素（外蛋白）在增强TSST-1跨表面上的渗透中的贡献，通过对上皮细胞的细胞溶解和/或炎症对上皮细胞的影响，以及2）确定TSST-1受体结合的机械构成，并引起tSST-1受体的启动。信号通路，随后的细胞因子产生。我们的初步数据表明，促进TSST-1在粘膜屏障中渗透的最可能的候选蛋白质是细胞毒素，包括3毒素，我们建议通过使用纯化的3毒素成分和等位基因替代敲除刺激曲折，以深入研究。 In the course of these studies, we will also study mucosal penetration and activities of a deletion [missing amino acids 1 to 72] mutant TSST-1 protein (13,000 MW vs. 22,000 MW wild-type TSST-1) that maintains superantigenicity and is associated with a rapidly progressive, 100% fatal, extreme pyrexia syndrome in humans.我们认为，计划的研究将阐明TSST-1穿透粘膜壁垒以引起大量人类疾病的机制。这些研究可能建议采用新的治疗策略来管理严重的金黄色葡萄球菌疾病。公共卫生相关性：细菌金黄色葡萄球菌经常从生物体的粘膜定植中引起严重的人类疾病，然后是分泌有效的外毒素，称为上皮。疾病包括可能具有月经和非月经形式的毒性休克综合征，在美国，每年可能会影响数千人类。该应用研究了超抗原渗透粘膜表面的机制，使金黄色葡萄球菌能够引起威胁生命的毒性休克综合症，并可能提出预防和管理此类疾病的新型策略。