Development of oligosaccharide conjugates that could be potentially utilized as vaccines against pathogenic Gram-negative bacteria

开发可潜在用作致病性革兰氏阴性菌疫苗的寡糖缀合物

基本信息

批准号：
14360207
负责人：
YAMASAKI Ryohei
金额：
$ 8.9万
依托单位：
Tottori University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

项目摘要

The object of this research is to study the possibility to develop carbohydrate vaccines t against pathogenic Gram-negative bacteria. We accomplished the following during the funding period.1)Synthesis of branched core oligosaccharide(OS) expressed in 15253 LOS and OS conjugates. 15253 We developed a new heptose (Hep) intermediate, 3-O-silyl-Hep, for the synthesis of the core OS. By utilizing the Hep intermediate, we constructed the following branched core OS : the 2,3-,3,4-branched, and 2,3;3,4-dibranched structures. We also developed a basic strategy to synthesize OS conjugates using a model OS. Finally, we synthesize a-spacer KDO side by stereoselective anomeric O-acylation.2)Analysis of antibodies specific for the core OS and the immunogenicity of a model OS conjugate. We characterized several antibodies that are specific for the core OS and showed that those core OS structures are immunogenic in humans for the first time, We also developed a sensitive western blot assay that would allow the detection of LOS (femt mol level) with lower concentration of antibodies and used this assay for the analysis of specificities of the above antibodies and anti-sera raised using the model OS conjugate.Thus, we established a methodology for the synthesis of branched core OS and have shown that such core OS can be utilized for the development of vaccines against pathogenic Gram-negative bacteria.

这项研究的目的是研究针对致病性革兰氏阴性细菌开发碳水化合物疫苗T的可能性。我们在资金期间完成了以下内容。1）合成在15253 LOS和OS共轭物中表达的分支核心寡糖（OS）。 15253我们开发了一种新的含素糖（HEP）中级3-O-Silyl-Hep，用于合成核心OS。通过利用HEP中间体，我们构建了以下分支的核心OS：2,3-，3,4分支和2,3; 3,4二元结构。我们还制定了一种基本策略，可以使用模型OS合成OS共轭物。最后，我们通过立体选择性异构O-acylation合成A-A-Spacer KDO侧。2）分析对核心OS特异的抗体和模型OS共轭物的免疫原性。我们表征了几种针对核心OS的抗体，并表明这些核心OS结构在人类中是第一次在人类中免疫原性，我们还开发了一种敏感的蛋白质印迹测定法，该测定允许检测LOS（femT MOL水平）具有较低浓度的抗体浓度，并将此测定法用于对上述抗体和抗体的特异性进行分析，我们使用了抗体的特异性。分支芯OS的合成，并表明可以将这种核心O用于开发针对致病性革兰氏阴性细菌的疫苗。