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The relevance between growth ability of Entamoeba dispar in tissue and its pathogenicity

迪斯帕内阿米巴在组织中的生长能力与其致病性的相关性

基本信息

批准号：
10670237
负责人：
KOBAYASHI Seiki
金额：
$ 1.15万
依托单位：
Keio University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670237/
关键词：
Entamoeba dispar Axenic culture Ferredoxin YIGADHA-S medium Growthability in tissue pathogenicity

项目摘要

We designed a new medium (YIGADHA-S) based on the casin-free YI-S medium (Diamond, 1995) and the monoxenic culture system with Pseudomonas aeruginosa (Kobayashi et al, 1998) for trail of axenic cultivation of Entamoeba dispar YIGADRA-S is deferent from YI-S in that glucose is replaced by gluconic acid and dihydroxyactone, and the medium is sterilized by filtration. Consequently this YIGADHA-S made successful axenic culture of four E. dispar strains, However the growth of E. dispar strains were very poor except for one strain, We recently found that various cell lines from lots of species (over 18 species) of animal or plant origin, which were autoclaved (121℃, 15min.), also could support the growth of E. dispar in addition to bacteria (Eschericia coli, P. aeruginosa and others) and Crithidia fasciculate, and found an interesting property common to every cell (e.g. micro organisms, protozoan, mammalian and plant cells) for supporting the growth of E. dispar in YIGADHA-S, That is, those … More cells except for bacteria had mitochondria, hydrogenosome or mitochondria and chroloplast. On the other hand, some protozoan (Entamoeba histolytica, E. dispar, E. moshkovskii (Laredo), E. invadens (IP-1), Ciardia lamblia (Portland I) cells and human erythrocyte which lacked mitochondria or mitochondria like organelles didn't make support the growth of E. dispar in this medium. We supposed from these results that the common nonheme iron-sulfur protein existing in mitochondria, hydrogenosome, chroloplast and bacteria might support the growth of E. dispar. And the effect of purified ferredoxin (Sigma) from spinach (ferredoxin is known as an essential redox protein in the energy metablism of E. histolytica) on the growth of E. dispar was tested. It was confirmed that ferredoxin (2.5 μg/ml【less than or equal】) had a favorable effect on the growth of E. dispar. And we developed a simple method for extraction of ferredoxin-rich fraction from isolated and washed intact leaf cells containing many chroloplasts of spiderwort (Commelina communis L.) by sonication. Consequently, this ferredoxin-rich extract made successful axenic culture of five E. dispar strains at similar maximum growth rate (constantly 3-4x10ィイD14ィエD1/ml). And' we also established an in vivo system for infection of E. dispar in caecum by using Rag gene knockout immunodeficient mouse (BALB/c Rag -/-), though the period of infection was for 3 days. Less

我们在无酪蛋白YI-S培养基（Diamond，1995）和铜绿假单胞菌单菌培养系统（小林et al，1998）的基础上设计了一种新的培养基（YIGADHA-S），用于舞毒蛾内阿米巴的无菌培养试验。结果表明，该YIGADHA-S菌株成功地对四株E. dispar菌株的生长，而E.我们最近发现，来自许多种（超过18种）动植物的各种细胞系，经高压灭菌（121℃，15分钟），也能支持E. dispar除了细菌（土杆菌、铜绿假单胞菌和其他）和束状短壳虫之外，还发现了每种细胞（例如微生物、原生动物、哺乳动物和植物细胞）共同的支持E.在YIGADHA-S dispar，即那些 ...更多信息除细菌外，其余细胞均具有线粒体、氢酶体或线粒体和叶绿体。另一方面，一些原生动物（溶组织内阿米巴，E. dispar、异枝野螟E. moshkovskii（拉雷多），E.侵袭菌（IP-1）、兰氏贾第鞭毛虫（波特兰I）细胞和缺乏线粒体或线粒体样细胞器的人红细胞不能支持E.在这个媒介中。推测线粒体、氢化酶体、叶绿体和细菌中存在的共同的非血红素铁硫蛋白可能支持大肠杆菌的生长。差异从菠菜中提取纯化的铁氧还蛋白（Sigma）（铁氧还蛋白是大肠杆菌能量代谢中必需的氧化还原蛋白），histolytica）对E.测试了Dispar。证实铁氧还蛋白（2.5 μg/ml[≤]）对E.差异本研究还建立了一种从鸭跖草（Commelina communis L.）通过超声处理。结果表明，该富含铁氧还蛋白的提取物成功地对5株大肠杆菌进行了纯培养。不同菌株在相似的最大生长速率下（恒定3- 4 × 10 ~（-1）D14 × 10 ~（-1）D1/ml）。同时，我们还建立了一个感染E.用Rag基因敲除的免疫缺陷小鼠（BALB/c Rag -/-）感染盲肠，感染期为3d。少