Bacillus anthracis cell surface carbohydrates

描述（由申请人提供）：关于炭疽芽孢杆菌的生物恐怖威胁要求开发改进的疫苗、诊断和治疗剂。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，对炭疽芽孢杆菌细胞壁的碳水化合物知之甚少。炭疽杆菌产生的荚膜由聚g-谷氨酸组成，而不是多糖。然而，最近的研究表明，在炭疽芽孢杆菌的营养和孢子形式中都有重要的碳水化合物成分。营养细胞壁含有一种多糖，它与肽聚糖相连，并通过与蛋白质的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。孢子的外孢子囊外壳含有一种主要的糖蛋白，它是孢子的免疫优势抗原。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芽孢杆菌特有的结构特征。没有关于胞外孢子糖蛋白碳水化合物特征的报道。据推测，炭疽芽胞杆菌细胞壁含有碳水化合物（例如s层锚定多糖和外孢糖蛋白），这些碳水化合物是物种和/或菌株特异性的，因此是开发疫苗、诊断和治疗剂的候选物。本应用程序的具体目的是：(a)确定s层锚定多糖和外生孢子碳水化合物是否具有物种特异性结构特征，(b)确定这些相同的碳水化合物是否具有菌株特异性结构特征，以及(c)确定炭疽芽孢杆菌毒力质粒pX01和pX02是否编码结构上修饰这些碳水化合物的酶。这项工作将通过从几种蜡样芽孢杆菌中分离和结构表征这些碳水化合物来完成，包括从几种菌株/分离的炭疽芽孢杆菌中分离出来的碳水化合物，以及从经pX01或pX02或两种质粒不同处理的炭疽芽孢杆菌的衍生物中分离出来的碳水化合物。这项工作将由乔治亚大学复合碳水化合物研究中心的罗素·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·奎因博士共同完成。

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.