PURIFICATION AND MECHANISM OF C. DIFFICILE TOXIN

艰难梭菌毒素的纯化及其机理

• 批准号: 3153220
• 负责人: JOHN THOMAS LAMONT
• 金额: $ 13.41万
• 依托单位: BOSTON UNIVERSITY MEDICAL CENTER HOSP
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-04-01 至 1987-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3153220
• 关键词: Clostridium; colitis; cytoskeleton; diarrhea; enterotoxins; epithelium; extracellular matrix; fibroblasts; high performance liquid chromatography; immunofluorescence technique; membrane permeability; phosphorylation; smooth muscle; synchronous cell division; ulcerative colitis

Clostridium difficile, the causative agent of antibiotic-associated diarrhea and colitis, is now recognized as one of the major enteric pathogens. C. difficile is an anaerobic bacillus which releases several protein toxins that mediate tissue damage in the colon of infected animals or humans. The major toxin, toxin B, is a heat labile protein which causes rounding of cells growing in monolayer culture. This toxin is also demonstrable in the stools of patients with antibiotic-associated diarrhea and colitis, and presumably mediates the tissue injury (colitis) which accompanies infection with this pathogen. The overall goals of this proposal are to purify C. difficile toxin B and determine its mechanism of action in vitro and in vivo. The hypothesis to be tested in this project is that toxin B interferes with the structure and function of the cytoskeleton, causing cell rounding, and that this in turn causes inhibition of macromolecular synthesis and eventually death of the cell. The purified toxin will be characterized as regards composition, molecular weight, sub-unit structure and structure-activity relationships. The purified toxin will then be used to determine the mechanism of cell rounding, more specificially its effects on cytoskeletal and matrix proteins. The syntehsis of actin, fibronectin, myosin and other filament proteins will be studied in fibroblast and smooth muscle cells exposed to graded doses of pure toxin B. The effect of toxin B on macromolecular synthesis will be quantitated using 3H-leucine and 3H-thymidine as precursors for protein and DNA syntehsis. The significance of these studies relates to the importance of C. difficile as a human pathogen. By providing a better understanding of how toxin B works it should be possible to define the pathophysiology of one of the commonest and most important enteric infections in man. From a basic science viewpoint, these studies should provide insights into the relationship of cytoskeleton and matrix proteins to cell shape, synthetic function and proliferation.

艰难梭菌，与抗生素相关的病原体

项目成果

Clostridium difficile cytotoxin inhibits protein synthesis in fibroblasts and intestinal mucosa.

艰难梭菌细胞毒素抑制成纤维细胞和肠粘膜的蛋白质合成。

• DOI: 10.1016/s0016-5085(86)80010-5
• 发表时间: 1986
• 期刊: Gastroenterology
• 影响因子: 29.4
• 作者: Pothoulakis,C;Triadafilopoulos,G;Clark,M;Franzblau,C;LaMont,JT
• 通讯作者: LaMont,JT

Purification and properties of Clostridium difficile cytotoxin B.

艰难梭菌细胞毒素 B 的纯化和性质。

• 发表时间: 1986
• 期刊: The Journal of biological chemistry
• 作者: Pothoulakis,C;Barone,LM;Ely,R;Faris,B;Clark,ME;Franzblau,C;LaMont,JT
• 通讯作者: LaMont,JT

Comparative study of Clostridium difficile toxin A and cholera toxin in rabbit ileum.

兔回肠中艰难梭菌毒素A与霍乱毒素的比较研究。

• DOI: 10.1016/0016-5085(89)91689-2
• 发表时间: 1989
• 期刊: Gastroenterology
• 影响因子: 29.4
• 作者: Triadafilopoulos,G;Pothoulakis,C;Weiss,R;Giampaolo,C;Lamont,JT
• 通讯作者: Lamont,JT

Differential effects of Clostridium difficile toxins A and B on rabbit ileum.

艰难梭菌毒素 A 和 B 对兔回肠的不同影响。

• DOI: 10.1016/0016-5085(87)91014-6
• 发表时间: 1987
• 期刊: Gastroenterology
• 影响因子: 29.4
• 作者: Triadafilopoulos,G;Pothoulakis,C;O'Brien,MJ;LaMont,JT
• 通讯作者: LaMont,JT

The chemotactic response of human granulocytes to Clostridium difficile toxin A is age dependent.

人粒细胞对艰难梭菌毒素 A 的趋化反应具有年龄依赖性。

• 发表时间: 1991
• 期刊: The American journal of gastroenterology
• 作者: Triadafilopoulos,G;Shah,MH;Pothoulakis,C
• 通讯作者: Pothoulakis,C