Bacillus anthracis cell surface carbohydrates

描述(由申请人提供)：与炭疽杆菌有关的生物恐怖威胁要求开发改进的疫苗、诊断和治疗试剂。细菌细胞表面的碳水化合物已经被用于所有这三个目的，然而，对炭疽杆菌细胞壁的碳水化合物知之甚少。炭疽杆菌产生的胶囊由多聚-g-谷氨酸组成，而不是多糖。然而，最近的研究表明，炭疽杆菌的营养形态和孢子形态中都有重要的碳水化合物成分。营养细胞壁含有一种与肽聚糖相连的多糖，通过与蛋白质的SLH结构域结合，将结晶表面(S层)蛋白质固定在细胞壁上。这种多糖被称为S层锚定多糖。孢子的外孢子层含有一种主要的糖蛋白，它是孢子的一种免疫优势抗原。虽然S层锚定多糖的结构尚未确定，但间接数据表明，它具有炭疽杆菌特有的结构特征。目前还没有关于外孢子菌糖蛋白的碳水化合物的报道。据推测，炭疽杆菌细胞壁含有碳水化合物(如S层锚定多糖和外孢子菌糖蛋白)，这些碳水化合物是物种和/或菌株特有的，因此是疫苗、诊断和治疗药物开发的候选物质。本申请的具体目的是(a.)确定S层锚定多糖和外孢子菌碳水化合物是否具有物种特有的结构特征，(b.)确定这些碳水化合物是否具有菌株特有的结构特征，以及(c.)确定炭疽杆菌毒力质粒pX01和pX02是否编码对这些碳水化合物进行结构修饰的酶。这项工作将通过从蜡样芽孢杆菌组的几个物种，包括炭疽杆菌变种Ames，从炭疽杆菌的几个菌株/分离物，以及从已经用pX01或pX02或两个质粒不同治愈的炭疽杆菌变种Ames衍生物中分离和结构特征来完成。这项工作将是佐治亚大学复合碳水化合物研究中心的拉塞尔·卡尔森博士、埃默里大学的大卫·斯蒂芬斯博士和疾病控制中心的康拉德·奎因博士共同努力的结果。

DESCRIPTION (provided by applicant): The bioterrorist threat with regard to Bacillus anthracis requires the development of improved vaccines, diagnostic, and therapeutic agents. Bacterial cell surface carbohydrates have been utilized for all three of these purposes, however, little is known about the B. anthracis cell wall carbohydrates. B. anthracis produces a capsule consisting of poly-g-glutamic acid, rather than a polysaccharide. However, recent work has shown that there are important carbohydrate components in both the vegetative and spore forms of B.anthracis. The vegetative cell wall contains a polysaccharide that is linked to peptidoglycan and serves to anchor the crystalline surface (S-layer) protein to the cell wall by binding with the SLH domain of the protein. This polysaccharide is referred to as the S-layer anchoring polysaccharide. The exosporium coat of the spore contains a major glycoprotein that is an immunodominant antigen of the spore. While the structure of the S-layer anchoring polysaccharide has not been determined, indirect data suggest that it has structural features that are specific to B. anthracis. There are no reports characterizing the carbohydrates of the exosporium glycoprotein. It is hypothesized that the B. anthracis cell wall contains carbohydrates (e.g. the S-layer anchoring polysaccharide and the exosporium glycoprotein) that are species and/or strain specific and are, therefore, candidates for the development of vaccines, diagnostic and therapeutic agents. The specific aims of this application are to (a.) determine if the S-layer anchoring polysaccharide and the exosporium carbohydrates have species specific structural features, (b.) determine if these same carbohydrates have strain specific structural features, and (c.) determine if the B. anthracis virulence plasmids, pX01 and pX02, encode for enzymes that structurally modify these carbohydrates. This work will be done by isolating and structurally characterizing these carbohydrates from several species of the B. cereus group, including B. anthracis var Ames, from several strains/isolates of B. anthracis, and from B.anthracis var Ames derivatives that have been differentially cured of pX01 or pX02, or of both plasmids. This work will be a collaborative effort between Dr. Russell Carlson at the Complex Carbohydrate Research Center at the University of Georgia, Dr. David Stephens at Emory University, and Dr. Conrad Quinn at the Centers for Disease Control.