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MECHANISM OF ACTION OF PORE FORMING TOXINS

成孔毒素的作用机制

基本信息

批准号：
06454206
负责人：
SHINODA Sumio
金额：
$ 4.61万
依托单位：
OKAYAMA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06454206/
关键词：
Pathogenic vibrios Hemolysin Osmotic hemolysis Vibrio mimicus Vibrio vulnificus Vibrio fluvialis PCR

项目摘要

Bacteria of genus Vibrio are normal habitant in natural environmental water. Because of universal consumption of sea foods in Japan, gastroenteric infections due to pathogenic vibrios break out frequently, therefore it is an important problem to clarify the mechanism of infection of the vibrios. Many pathogenic vibrios produce hemolysin as an important pathogenic factor. It is expected that elucidation of action mechanism of the hemolysins leads to development of methods for prevention and therapy of the vibrio infections. We selected two vibrio hemolysins, Vibrio vulnificus hemolysin (VVH) and V.mimicus hemolysin (VMH), and carried out the comparative study of action mechanism in vitro and in vivo. The project started in 1994. Differences of properties of the hemolysins produced by different strains and a role of VMH as a diarrhenogenic factor were demonstrated in 1994 and 1995. In 1996, protein-chemical and immuno-chemical studies of VMH,especially structure and function, were carried out. VMH was purified with ethylene glycol as the solvent. Monoclonal antibodies (MAb) against VMH were prepared but only IgM MAbs which had binding activity to VMH but not neutralizing activity were obtained. Furthermore, non-immunized IgMs also showed weak binding activity to VMH which was suspected to bind to sialic acid on the IgM molecule. The IgM antibodies, however, showed neutralizing activity to VMH purified by a modified method without ethylene glycol, suggesting the conformational change of recognition site for sialic acid of the membrane surface by ethylene glycol, although it is not so distinguish detectable by physico-chemical methods such as fluorescence analysis. Informations on mechanism of binding of VMH molecule to target cells obtained in this project are useful for understanding of the mechanism of gastroenteritis due to the vibrio.

弧菌属细菌是自然环境水体中常见的细菌。在日本，由于海产品的普遍消费，由致病性弧菌引起的胃肠道感染频繁爆发，因此阐明弧菌的感染机制是一个重要的问题。溶血素是许多致病性弧菌产生的重要致病因子。人们期望阐明溶血素的作用机制，从而开发预防和治疗弧菌感染的方法。本研究选取了两种弧菌溶血素--创伤弧菌溶血素（VVH）和拟态弧菌溶血素（VMH），并对其作用机制进行了体内外比较研究。该项目始于1994年。1994年和1995年证明了不同菌株产生的溶血素性质的差异以及VMH作为一种致溶血因子的作用。1996年，人们对VMH进行了蛋白质化学和免疫化学研究，特别是结构和功能的研究。用乙二醇作为溶剂纯化VMH。制备了抗VMH的单克隆抗体（MAb），但仅获得了具有结合VMH活性而没有中和活性的IgM MAb。此外，未免疫的IgM也显示出对VMH的弱结合活性，VMH被怀疑与IgM分子上的唾液酸结合。然而，IgM抗体显示出对VMH的中和活性，该VMH通过不含乙二醇的改进方法纯化，这表明乙二醇对膜表面的唾液酸识别位点的构象变化，尽管通过物理化学方法如荧光分析不能如此区分检测。本研究所获得的VMH分子与靶细胞结合机制的信息，对于理解弧菌引起胃肠炎的机制具有重要意义。