Bacillus anthracis cell surface carbohydrates

描述（由申请人提供）：炭疽杆菌的生物恐怖威胁需要开发改进的疫苗、诊断和治疗剂。 细菌细胞表面碳水化合物已被用于所有这三个目的，然而，很少有人知道的B。炭疽菌细胞壁碳水化合物。 B。炭疽菌产生由聚-g-谷氨酸而不是多糖组成的胶囊。 然而，最近的研究表明，有重要的碳水化合物成分在营养和孢子形式的炭疽杆菌。 营养细胞壁含有多糖，该多糖与肽聚糖连接，并通过与蛋白质的SLH结构域结合而将结晶表面（S层）蛋白质锚到细胞壁。 这种多糖被称为S层锚定多糖。 孢子的外孢壁外壳含有一种主要的糖蛋白，它是孢子的免疫显性抗原。 虽然尚未确定S层锚定多糖的结构，但间接数据表明其具有B特异性的结构特征。炭疽病 没有报道表征外孢壁糖蛋白的碳水化合物。 假设B.炭疽菌细胞壁含有碳水化合物（例如S-层锚定多糖和外孢壁糖蛋白），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、诊断剂和治疗剂的候选物。 本申请的具体目的是（a.）确定所述S层锚定多糖和所述孢子外壁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B.炭疽毒力质粒pX 01和pX 02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B中分离和结构表征这些碳水化合物来完成。蜡状菌群，包括B.炭疽变种艾姆斯，来自B的几个菌株/分离株。炭疽杆菌，和来自已被pX 01或pX 02或两种质粒差异治愈的B.anthracis var艾姆斯衍生物。 这项工作将是格鲁吉亚大学复杂碳水化合物研究中心的Russell Carlson博士、埃默里大学的大卫斯蒂芬斯博士和疾病控制中心的Conrad Quinn博士之间的合作努力。

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.