CARDIOTOXICITY OF STREPTOCOCCAL PYROGENIC EXOTOXINS

链球菌热原性外毒素的心脏毒性

• 批准号: 6183079
• 负责人: Patrick M Schlievert
• 金额: $ 17.14万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-07-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6183079
• 关键词: Staphylococcus; Streptococcus; T cell receptor; cardiotoxin; drug delivery systems; enterotoxins; epitope mapping; exotoxins; laboratory rabbit; lipopolysaccharides; molecular pathology; nucleic acid sequence; protein purification; protein sequence; protein structure function; pyrogens; site directed mutagenesis; superantigens

DESCRIPTION (Adapted from the applicant's abstract): The long term goals of this project are two fold: a) to evaluate the role of pyrogenic toxin superantigens, notably streptococcal pyrogenic exotoxins (SPEs, scarlet fever toxins), in causing both acute toxic shock syndrome and vascular illnesses and chronic autoimmune and allergic diseases, and b) to analyze the structure-function relationships among the SPEs and between the SPEs and staphylococcal enterotoxins and toxic shock syndrome toxin-1, with the intent to clarify the molecular mechanism(s) of action of the toxins and develop toxoid vaccines against the toxins. Specific aims of the present application include: a) determination of the three dimensional structure of SPE C and SPEA/staphylococcal enterotoxin C (SEC/SEB complexed. with the T cell receptor beta chain. The investigator's role in these studies is to provide sufficient toxins for structural analyses by ethanol precipitation from culture fluids, resolubilization in acetate buffered saline at pH 4.0 or water, and preparative isoelectric focusing. Crystallization and three dimensional structure analysis of SPE C will be done in collaboration with Dr. Douglas H. Ohlendorf, Department of Biochemistry, University of Minnesota and that of SPE A/SEB complexed to the beta chain of the T cell receptor by Dr. Roy A.. Mariuzza, Center for Advanced Research in Biotechnology, University of Maryland; b) Domains and amino acid residues on SPE C and the SPE A/SEC/SEB subgroup of pyrogenic toxin superantigens required for biological activity (pyrogenicity, enhancement of lethal endotoxin shock and cardiotoxicity, ability to induce TSS when administered subcutaneously in miniosmotic pumps, superantigenicity, and lipopolysaccharide binding) will be localized through use of PCR mutagenesis. Nucleotide sequencing will be done to verify changed amino acids and structural analysis where possible to assess alterations of three dimensional structure of mutants. It is hoped in addition to localizing domains required for toxicity, that these studies will clarify important mechanisms of T cell activation and lead to useful toxoid vaccines.

描述（改编自申请人的摘要）：长期目标 该项目有两个方面：a）评估热原毒素的作用 超抗原，特别是链球菌致热外毒素（SPE，猩红 发热毒素），在引起急性中毒性休克综合征和血管 疾病和慢性自身免疫性和过敏性疾病，和B）分析 SPE之间以及SPE与 葡萄球菌肠毒素和中毒性休克综合征毒素-1， 旨在阐明毒素作用的分子机制， 开发针对毒素的类毒素疫苗。 本申请的具体目的包括： SPE C和SPEA/葡萄球菌肠毒素C的三维结构 (SEC/SEB复合。 与T细胞受体β链连接 的 研究人员在这些研究中的作用是提供足够的毒素， 通过从培养液中乙醇沉淀进行结构分析， 在pH 4.0的乙酸盐缓冲盐水或水中再溶解，和 制备等电聚焦 结晶与三维 SPE C的结构分析将与道格拉斯博士合作进行 H. Ohlendorf，明尼苏达大学生物化学系， SPE A/SEB与T细胞受体的β链复合， A.. Mariuzza，生物技术高级研究中心， 马里兰州; B）SPE C和SPE A/SEC/SE B上的结构域和氨基酸残基 生物活性所需热原毒素超抗原亚群 （致热原性、致死性内毒素休克和心脏毒性的增强， 在微渗透泵中皮下给药时诱导TSS的能力， 超抗原性和脂多糖结合）将通过 使用PCR诱变。 将进行核苷酸测序以验证 改变氨基酸和结构分析（如可能），以评估 突变体三维结构的改变。 希望在 除了定位毒性所需的结构域，这些研究 将阐明T细胞活化的重要机制，并导致有用的 类毒素疫苗