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Research on pathogenic gienes and development of vaccine of Vibrio valunificus, a novel causative agent of foodborne infection

食源性感染新病原瓦卢弧菌病原学研究及疫苗研制

基本信息

批准号：
22590400
负责人：
YAMAMOTO Kouichirou
金额：
$ 2.91万
依托单位：
Okayama Prefectural University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2010
资助国家：
日本
起止时间：
2010 至 2012
项目状态：
已结题

项目摘要

Vibrio vulnificus is an important pathogens of foodborn diseases and wound infection. This pathogen causes fatal septicemia rapidly in immunocompromised hosts after they contact with contaminated seawater or seafood with this organism. The mortality rate is more than 50%. For the rapid progress and the high mortality,vaccination is considered an effective way to control infection with V. vulnificus in a high risk population. We applied Signature-Tagged Mutagenesis (STM) to generate attenuated mutants from a clinically-isolated virulent V. vulnificus strain, OPU1 which was isolated from a patient. Twelve insertion mutants were selected and the virulence of these mutants were determined by the minimal lethal dose (MLD) to mice. Several mutant strains showed lower virulence compare to the parent virulent strain. The DNA sequences flanking to transposon insertion sites in these mutants were determined and searched for homologies at DNA Date Bank of Japan. The transposons inserted genes were those for IMP dehydrogenase, UDP-N-acetylgulcosamin-2-epimerase, aspartokinase, and a conserved hypothetical protein. This suggests that these genes may be involved in V. vulnificusvirulence. One of the mutants was administrated for experimental vaccination to mice. The immunized mice were challenged with the virulent parent strain. All unimmunized mice were killed within 36 hr, but almost all immunized mice survived. In this experiment, we found that the attenuated have a potentiality as live vaccine candidate.

创伤弧菌是食源性疾病和伤口感染的重要病原菌。这种病原体在免疫功能低下的宿主接触受污染的海水或海产品后迅速引起致命的败血症。死亡率超过50%。由于创伤弧菌进展快、死亡率高，疫苗接种被认为是控制高危人群创伤弧菌感染的有效方法。我们应用签名标记诱变（STM）从临床分离的创伤弧菌强毒株OPU 1中产生减毒突变体，OPU 1是从患者中分离的。筛选出12个插入突变体，并通过小鼠最小致死量（MLD）测定这些突变体的毒力。几个突变株的毒力比亲本菌株低。确定这些突变体中转座子插入位点侧翼的DNA序列，并在日本DNA数据库中搜索同源性。转座子插入的基因是IMP脱氢酶、UDP-N-乙酰谷氨酰胺-2-差向异构酶、葡萄糖激酶和保守的假设蛋白。这表明这些基因可能参与创伤弧菌的毒力。将其中一种突变体用于小鼠的实验性疫苗接种。用强毒株攻击免疫小鼠。所有未免疫的小鼠在36小时内被杀死，但几乎所有免疫的小鼠存活。本实验发现，该弱毒疫苗具有作为活疫苗候选物的潜力。

项目成果

期刊论文数量（0）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Vibrios as a causative agent of foodborn diseases.

弧菌是食源性疾病的病原体。

DOI：
发表时间：
2010
期刊：
影响因子：
0
作者：
Yamamoto;K.
通讯作者：
K.

Vibrio vulnficus溶血性に関する遺伝子

创伤弧菌溶血相关基因

DOI：
发表时间：
2012
期刊：
影响因子：
0
作者：
山本耕一郎;他4名
通讯作者：
他4名

Vibrio vulnificus トランスポゾン挿入弱毒化変異株と挿入部位遺伝子の特定

创伤弧菌转座子插入减毒突变株及插入位点基因的鉴定

DOI：
发表时间：
2010
期刊：
影响因子：
0
作者：
Nakata;K.;Yamamoto;M.;Yamamoto;Inagawa;H.;Soma;G;山本真衣,北林奏,肖建波,瀬川理恵,井上美幸,細原浩平,横田憲治,小熊恵二,山本耕一郎
通讯作者：
山本真衣,北林奏,肖建波,瀬川理恵,井上美幸,細原浩平,横田憲治,小熊恵二,山本耕一郎

Molecular structure-affinity relationship of natural polyphenols for bovine・-globulin

天然多酚对牛球蛋白的分子结构-亲和关系