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.

本研究的目的是研究开发针对致病性革兰氏阴性菌的碳水化合物疫苗的可能性。在资助期间我们完成了以下工作：1）以15253个LOS和OS缀合物表达的支链核心寡糖（OS）的合成。 15253 我们开发了一种新的庚糖 (Hep) 中间体 3-O-甲硅烷基-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核心操作系统：2,3-,3,4-分支和 2,3;3,4-二分支结构。我们还开发了一种使用模型 OS 合成 OS 缀合物的基本策略。最后，我们通过立体选择性异头O-酰化合成了a-间隔基KDO侧。2)核心OS特异性抗体和模型OS缀合物的免疫原性分析。我们鉴定了几种针对核心操作系统的特异性抗体，并首次证明这些核心操作系统结构在人类中具有免疫原性。我们还开发了一种灵敏的蛋白质印迹测定法，可以检测较低浓度的 LOS（femt mol 水平）抗体并使用该测定法分析上述抗体和使用模型 OS 缀合物产生的抗血清的特异性。因此，我们建立了分支核心 OS 的合成方法，并表明这种核心 OS 可用于开发针对病原体的疫苗革兰氏阴性细菌。