CARDIOTOXICITY OF STREPTOCOCCAL PYROGENIC EXOTOXINS

描述(改编自申请人的摘要)：长期目标 本项目有两个方面：a)评估热原毒素的作用 超抗原，特别是链球菌热原外毒素(SPE、猩红 发热毒素)，导致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，以及b)分析 SPE之间以及SPE和SPE之间的结构-功能关系 葡萄球菌肠毒素和中毒性休克综合征毒素-1，与 意在阐明毒素作用的分子机制(S)和 开发针对毒素的毒素疫苗。 本申请的具体目的包括：a)确定 SpE C和SpEA/葡萄球菌肠毒素C的三维结构 (SEC/SEB复杂。与T细胞受体β链结合。这个 调查者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值为4.0的醋酸盐缓冲盐水或水中溶解，以及 制备性等电聚焦。结晶与三维 SPE C的结构分析将与道格拉斯博士合作完成 H·奥伦多夫，明尼苏达大学生物化学系 Roy博士将SpE A/SEB与T细胞受体的β链复合 一个.。马里乌扎，加州大学生物技术高级研究中心 马里兰州；b)SPE C和SPE A/SEC/SEB上的结构域和氨基酸残基 生物活性所需的热原毒素超抗原亚群 (致热性，增强致死性内毒素休克和心脏毒性， 在微渗泵中皮下给药时诱发TSS的能力， 超抗原性和脂多糖结合)将通过 利用聚合酶链式反应进行诱变。将进行核苷酸测序以验证 尽可能用改变的氨基酸和结构分析来评估 突变体的三维结构的改变。它被寄予希望 除了定位毒性所需的结构域外，这些研究 将阐明T细胞激活的重要机制，并导致有用的 类毒素疫苗。

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.