Estudios clínicos (inglés)

 


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SACCHAROMYCES BOULARDII PREVENTS TNF–INDUCED APOPTOSIS IN EHEC-INFECTED T84 CELLS

Guillaume Dalmassoa, Agnès Loubatb, Stéphanie Dahanc, Gustavo Called, Patrick Rampald and Dorota Czeruckaa,

Laboratoire de Gastroentérologie et Nutrition, IFR50, Faculté de Médecine, Université de Nice-Sophia Antipolis, 06107 Nice cedex 2, France

IFR50, Faculté de Médecine, Université de Nice-Sophia Antipolis, 06107 Nice cedex 2, France

Immunolobiology Center, 1 Gustave L. Levy, Place box 1089, 1029 New York, USA

Centre Hospitalier Princesse Grace, Service d’Hépato-Gastro-Entérologie, Avenue Pasteur BP 489, 92018 Monaco cedex

Received 29 May 2005;  accepted 3 November 2005.  Available online 13 January 2006.

doi:10.1016/j.resmic.2005.11.007

Copyright © 2006 Elsevier SAS All rights reserved.

Abstract

Induction of apoptosis and necrosis by enterohemorrhagic Escherichia coli (EHEC) has been reported in vivo and in vitro, but features of cell death were not noted in those reports. Since tumor necrosis factor-alpha (TNF-) has been implicated in the apoptosis of invasive bacteria, we investigated the role of this cytokine in EHEC-induced apoptosis. We hypothesize that the probiotic yeast strain Saccharomyces boulardii that interferes with EHEC-induced pro-inflammatory pathways delays EHEC-induced apoptosis. By 6 h of infection, flow cytometry analysis of T84 cells demonstrated that 40% of cells were FITC–annexin-V-positive and 40% of cells incorporated both annexin and propidium iodide (PI). Simultaneously, western blot analysis demonstrated that procaspases-8 and -3 were cleaved. Fragmentation of internucleosomal DNA revealed evidence of apoptotic leader formation after 8 and 9 h of infection. Procaspase-9 activation and 3′,3-dihexyloxacarbocyanine iodide (DiOC6) incorporation were observed at 3 h of infection. In cells preincubated with S. boulardii and infected with EHEC in the presence of yeast, the quantities of procaspases-8, -9 and -3 did not vary, and no DNA fragmentation was observed. The TNF- transcript level and the level of secreted TNF- increased considerably (P<0.001 vs control cells) at 6 h of infection in EHEC-alone-infected cells, but were significantly reduced in cells infected in the presence of S. boulardii (P<0.001 vs EHEC-alone-infected cells). The presence of anti-TNF- antibody during infection reduced by 30% the level of FITC–annexin V-positive cells. Altogether, these findings demonstrated that: (i) EHEC infection stimulated TNF- synthesis that is implicated in apoptosis of T84 cells; and (ii) S. boulardii induced a decrease in TNF- and related apoptosis in EHEC-infected T84 cells. Keywords: EHEC; TNF-; Saccharomyces boulardii; Probiotic; Apoptosis Adherence of Escherichia coli serogroup O 157 and the Salmonella typhimurium mutant DT 104 to the surface of Saccharomyces boulardii. Gedek BR. Sachverständige für Mikroökologie und Mykotoxinologie, Ismaning, Germany. The detection of lectin sites for mannose-sensitive adhesion in the outer membrane of Saccharomyces boulardii and the irreversible binding of both enteropathogenic Escherichia coli (EPEC) and salmonellae (serovar Salmonella Typhimurium and Salmonella Enteritidis) provided the motivation to carry out further investigations to find out whether also other enteric bacteria such as entero-haemorrhagic Escherichia coli (EHEC) and the DT 104 mutant of S. Typhimurium have the capacity for binding to the cell wall of this yeast. Reference strains and fresh isolates from clinical cases of EHEC infections as well as salmonellae of the DT 104 mutant were included in this study using the agglutination test. The results first of all showed that EHEC of the serogroup O 157 and the DT 104 mutant of S. Typhimurium were bound to the surface of Saccharomyces boulardii. Because these bacteria do not respond very well to drugs but most of the gastrointestinal infections are caused by them, the use of S. boulardii for treatment and prophylaxis could be an excellent alternative. PMID: 10424093 [PubMed – indexed for MEDLINE]


SACCHAROMYCES BOULARDII STIMULATES INTESTINAL IMMUNOGLOBULIN A

Immune Response to Clostridium difficile Toxin A in Mice

INFECTION AND IMMUNITY,

0019-9567/01/$04.0010 DOI: 10.1128/IAI.69.4.2762–2765.2001

Apr. 2001, p. 2762–2765 Vol. 69, No. 4

Copyright © 2001, American Society for Microbiology. All Rights Reserved.

AMIR QAMAR, SAMER ABOUDOLA, MICHEL WARNY, PIERRE MICHETTI, CHARALABOS POTHOULAKIS, J. THOMAS LAMONT, AND CIARA´N P. KELLY*

Gastroenterology Division, Beth Israel Deaconess Medical Center,

Harvard Medical School, Boston, Massachusetts 02215

Received 26 October 2000/Returned for modification 13 December 2000/Accepted 16 January 2001

Saccharomyces boulardii is a nonpathogenic yeast that protects against antibiotic-associated diarrhea and recurrent Clostridium difficile colitis. The administration of C. difficile toxoid A by gavage to S. boulardii-fed BALB/c mice caused a 1.8-fold increase in total small intestinal immunoglobulin A levels (P 5 0.003) and a 4.4-fold increase in specific intestinal anti-toxin A levels (P < 0.001). Enhancing host intestinal immune responses may be an important mechanism for S. boulardii-mediated protection against diarrheal illnesses.


RESPONSE TO CHOLERA TOXIN OF 2 EPITHELIAL INTESTINAL CELL LINES. EFFECT OF SACCHAROMYCES BOULARDII

Czerucka D, Nano JL, Bernasconi P, Rampal P.
Laboratoire de Gastroentérologie et Nutrition, Faculté de Médecine,Nice.

Cholera toxin acts in vivo by activating intestinal adenylate cyclase. This study was designed to determine (1) whether normal rat epithelial intestinal cell lines (IRD 98 and IEC 17) respond to cholera toxin (CT) by an increased concentration of cyclic AMP and (2) whether the yeast Saccharomyces boulardii, which reduced CT-induced secretion of water and electrolytes using the isolated jejunal loop technique, has an effect on these models. The cAMP concentration evaluated in cells exposed to Saccharomyces boulardii and to cholera toxin (1 microgram/ml for 90 min) was compared to the concentration of cAMP obtained in control cells without yeast. Prior exposure of IRD 98 and IEC 17 cells to Saccharomyces boulardii, reduced CT-induced cAMP by 50 p. 100. This effect disappeared after destruction of the yeast by heating. Results show that the IRD 98 and IEC 17 cells are good models for in vitro investigation of the effects of cholera toxin. Our results suggests that Saccharomyces boulardii prevents the water and electrolyte secretion induced by cholera toxin.

PMID: 2544474 [PubMed – indexed for MEDLINE]


TREATMENT OF RECURRENT CLOSTRIDIUM DIFFICILE COLITIS WITH VANCOMYCIN AND SACCHAROMYCES BOULARDII

Surawicz CM, McFarland LV, Elmer G, Chinn J.

Department of Medicine, University of Washington, Seattle.

Recurrence of Clostridium difficile-associated diarrhea and pseudomembranous colitis occurs in up to 20% of patients after standard therapy. In these patients, subsequent recurrences are even more frequent. Saccharomyces boulardii, a nonpathogenic yeast, was found to be effective in preventing clindamycin cecitis recurrence in an animal model. We performed an open trial of S. boulardii to evaluate its efficacy in treating recurrences of C. difficile-associated colitis in humans. Thirteen patients with recurring C. difficile cytotoxin-positive diarrhea (who had an average of 3.6 previous recurrences) were treated with 10 days of vancomycin and a 30-day course of S. boulardii. Eleven (85%) had no further recurrences. S. boulardii may have a role in treating recurrent C. difficile diarrhea and colitis.

PMID: 2679049 [PubMed – indexed for MEDLINE]