Ulcerative Colitis: Swidsinski A

Swidsinski A, Loening-Baucke V, Bengmark S, Scholze J, Doerffel Y. · Humboldt University, Charité Hospital, CCM, Laboratory for Molecular Genetics, Polymicrobial Infections and Biofilms, Berlin, Germany. · Arch Med Res. · Pubmed #18164963 No free full text.

Abstract: BACKGROUND: Antibiotics are commonly used in inflammatory bowel disease (IBD). Little is known about their effect on the mucosal flora. METHODS: The mucosal flora was investigated in colonoscopic biopsies from six groups of 20 IBD patients each. Patients were selected with regard to duration of/interval to combined metronidazole and ciprofloxacin therapy: group I patients with 1 day and group II with 7-14 days of antibiotic therapy, group III-V patients evaluated 1-4 weeks, 2-18 weeks, 26-36 weeks after cessation of antibiotic therapy, respectively. The control group VI included patients without antibiotic therapy. Thirty different fluorescent in situ hybridization (FISH) probes representative of the diversity of the human intestinal flora were applied to all specimens. RESULTS: Bacteria adherent to mucosa could be seen exclusively in DAPI stain and were practically nonamenable to FISH probes in patients on antibiotics (0.001-3+/-0.001-5)x10(10)/mL. Occurrence and concentrations were significantly reduced in groups I and II as compared to untreated controls. The mucosal bacteria were significantly augmented after cessation of antibiotic therapy in group III (13.2+/-4.3) and group IV (5.8+/-2) but not in group V (1.1+/-0.8) as compared to group VI (0.5+/-0.4)x10(10)/mL. Neither Bacteroides nor Enterobacteriaceae groups were permanently suppressed by metronidazole-ciprofloxacin therapy. CONCLUSIONS: The suppressing effects of antibiotics on the mucosal flora are accompanied by massive rebound effects. The concentrations of mucosal bacteria are dramatically increased as soon as 1 week after cessation of antibiotic therapy, remaining at a level that is at least one power higher over a period of 5 months as compared to the group without antibiotic treatment.

Swidsinski A, Loening-Baucke V, Vaneechoutte M, Doerffel Y. · Humboldt University, Charité Hospital, Laboratory for Molecular Genetics, Polymicrobial Infections and Bacterial Biofilms, Berlin, Germany. · Inflamm Bowel Dis. · Pubmed #18050295 links to  free full text

Abstract: BACKGROUND: The intestinal microflora is important in the pathogenesis of inflammatory bowel disease (IBD). The impact of its spatial organization on health and disease is unknown. METHODS: We investigated sections of paraffin-embedded punched fecal cylinders. Fluctuations in spatial distribution of 11 bacterial groups were monitored in healthy subjects (n = 32), patients with IBD (n = 204), and other gastrointestinal diseases (n = 186) using fluorescence in situ hybridization (FISH). RESULTS: The microbial structure differed in patients with Crohn's disease (CD), ulcerative colitis (UC), and healthy and disease controls. The profiles of CD and UC were distinctly opposite in 6 of 11 FISH probes used. Most prominent were a depletion of Faecalibacterium prausnitzii (Fprau<1 x 10(9)/mL) with a normal leukocyte count in CD and a massive increase of leukocytes in the fecal-mucus transition zone (>30 leukocytes/10(4) microm(2)) with high Fprau in patients with UC. These 2 features alone enabled the recognition of active CD (Crohn's Disease Activity Index [CDAI] >150) or UC (Clinical Activity Index [CAI] >3) with 79%/80% sensitivity and 98%/100% specificity. The mismatch in the sensitivity was mainly due to overlap between single IBD entities, and the specificity was exclusively due to the similarity of Crohn's and celiac disease. When inflammatory bowel disease (IBD) patients were pooled the sensitivity was 100% for severe disease, 84% for moderate activity, 72% for IBD with < or =12 months remission, and 24% for IBD with >12 months remission. CONCLUSIONS: The fecal flora is highly structured and spatially organized. Diagnosing IBD and monitoring disease activity can be performed based on analysis of punched fecal cylinders independent from the patient's complaints.

Swidsinski A, Loening-Baucke V, Bengmark S, Lochs H, Dörffel Y. · Humboldt University, Charité Hospital, CCM, Laboratory for Molecular Genetics, Polymicrobial Infections and Bacterial Biofilms, 10098 Berlin, Germany. · Inflamm Bowel Dis. · Pubmed #17206639 links to  free full text

Abstract: BACKGROUND: The impact of azathioprine and 5-aminosalicylic acid (5-ASA) on the innate immunity and mucosal flora is unknown. The study investigated the influence of IBD treatment on the concentrations and spatial organization of mucosal bacteria using fluorescence in situ hybridization with 16s r-RNA targeting probes. METHODS: We prospectively investigated colonoscopic biopsies from five groups of 20 subjects each: patients with ulcerative or indeterminate colitis treated with azathioprine (group 1), azathioprine and 5-ASA (group 2), 5-ASA (group 3), untreated IBD (group 4), and healthy controls. RESULTS: The elevated numbers of leukocytes in mucus of IBD patients were reduced nearly to norm in patients treated with azathioprine alone. In contrast, 5-ASA therapy had no influence on mucus leukocyte migration and was associated with the lowest concentrations of mucosal bacteria of all IBD groups. The suppressed migration of leukocytes in azathioprine-treated patients was accompanied by a 28-fold higher concentration of mucosal bacteria when compared with the 5-ASA group or a 1000-fold increase when compared with healthy controls. The percent of the epithelial surface covered with adherent bacteria (P < 0.001) and the amenability of mucosal bacteria (P = 0.01) were also significantly increased in the azathioprine-treated group compared with all other IBD groups. The patients receiving both 5-ASA and azathioprine did not differ statistically from untreated IBD patients either in mucus leukocyte migration or in bacterial concentrations. CONCLUSIONS: Azathioprine and 5-ASA induce opposite effects on the mucus barrier. Concomitant therapy of 5-ASA and azathioprine mutually neutralizes the effects of both on the mucosal flora and the barrier function.

Swidsinski A, Loening-Baucke V, Theissig F, Engelhardt H, Bengmark S, Koch S, Lochs H, Dörffel Y. · Humboldt University, Charité, CCM, 10098 Berlin, Germany. · Gut. · Pubmed #16908512 No free full text.

Abstract: AIM: To study the role of mucus in the spatial separation of intestinal bacteria from mucosa. PATIENTS AND METHODS: Mucus barrier characteristics were evaluated using histological material obtained by biopsy from purged colon, colon prepared with enema and material from untreated appendices fixed with non-aqueous Carnoy solution. Bacteria were evaluated using fluorescence in situ hybridization, with bacterial 16S RNA probes and related to the periodic acid Schiff alcian blue stain. Biopsies from controls (n = 20), patients with self-limiting colitis (SLC; n = 20), ulcerative colitis (n = 20) and 60 randomly selected appendices were investigated. RESULTS: The mucosal surface beneath the mucus layer was free of bacteria in > or =80% of the normal appendices and biopsies from controls. The thickness of the mucus layer and its spread decreased with increasing severity of the inflammation; the epithelial surface showed bacterial adherence, epithelial tissue defects and deep mucosal infiltration with bacteria and leucocytes. Bacteria and leucocytes were found within mucus in all biopsy specimens from patients with ulcerative colitis, SLC, and acute appendicitis. The concentration of bacteria within mucus was inversely correlated to the numbers of leucocytes. CONCLUSIONS: The large bowel mucus layer effectively prevents contact between the highly concentrated luminal bacteria and the epithelial cells in all parts of the normal colon. Colonic inflammation is always accompanied by breaks in the mucus barrier. Although the inflammatory response gradually reduces the number of bacteria in mucus and faeces, the inflammation itself is not capable of preventing bacterial migration, adherence to and invasion of the mucosa.

Swidsinski A, Weber J, Loening-Baucke V, Hale LP, Lochs H. · Innere Klinik, Gastroenterologie, Charité Humboldt Universität, 10098 Berlin, Germany. · J Clin Microbiol. · Pubmed #16000463 links to  free full text

Abstract: The composition and spatial organization of the mucosal flora in biopsy specimens from patients with inflammatory bowel disease (IBD; either Crohn's disease or ulcerative colitis), self-limiting colitis, irritable-bowel syndrome (IBS), and healthy controls were investigated by using a broad range of fluorescent bacterial group-specific rRNA-targeted oligonucleotide probes. Each group included 20 subjects. Ten patients who had IBD and who were being treated with antibiotics were also studied. Use of nonaqueous Carnoy fixative to preserve the mucus layer was crucial for detection of bacteria adherent to the mucosal surface (mucosal bacteria). No biofilm was detectable in formalin-fixed biopsy specimens. Mucosal bacteria were found at concentrations greater than 10(9)/ml in 90 to 95% of IBD patients, 95% of patients with self-limiting colitis, 65% of IBS patients, and 35% of healthy controls. The mean density of the mucosal biofilm was 2 powers higher in IBD patients than in patients with IBS or controls, and bacteria were mostly adherent. Bacteroides fragilis was responsible for >60% of the biofilm mass in patients with IBD but for only 30% of the biofilm mass in patients with self-limiting colitis and <15% of the biofilm mass in patients with IBS. In contrast, bacteria which positively hybridized with the probe specific for Eubacterium rectale-Clostridium coccoides accounted for >40% of the biofilm in IBS patients but for <15% of the biofilm in IBD patients. In patients treated with (5-ASA) or antibiotics, the biofilm could be detected with 4,6-diamidino-2-phenylindole but did not hybridize with fluorescence in situ hybridization probes. A Bacteroides fragilis biofilm is the main feature of IBD. This was not previously recognized due to a lack of appropriate tissue fixation. Both 5-ASA and antibiotics suppress but do not eliminate the adherent biofilm.

Darfeuille-Michaud A, Boudeau J, Bulois P, Neut C, Glasser AL, Barnich N, Bringer MA, Swidsinski A, Beaugerie L, Colombel JF. · Pathogénie Bactérienne Intestinale, Laboratoire de Bactériologie, Université d'Auvergne, Clermont-Ferrand, France. · Gastroenterology. · Pubmed #15300573 No free full text.

Abstract: BACKGROUND & AIMS: Adherent-invasive Escherichia coli (AIEC) pathovar has been identified in the intestinal mucosa of patients with Crohn's disease (CD). AIEC reference strain LF82 is able to adhere to intestinal epithelial cells, to invade epithelial cells via a mechanism involving actin polymerization and microtubules, and to survive and replicate within macrophages. This study was performed to assess the prevalence of AIEC associated with intestinal mucosa of patients with CD, ulcerative colitis (UC), and of controls. METHODS: A search for E. coli strains was performed with ileal specimens of 63 patients with CD and 16 controls without inflammatory bowel disease (IBD), and with colonic specimens of 27 patients with CD, 8 patients with UC, and 102 controls. The abilities of E. coli strains to invade epithelial cells and to survive and replicate within macrophages were assessed using the gentamicin protection assay. Bacterial uptake by epithelial cells was analyzed using cytoskeletal inhibitors. Bacterial adhesion was quantified with Caco-2 and Intestine-407 cells. The presence of known E. coli virulence genes was assessed by polymerase chain reaction and DNA hybridization. RESULTS: In ileal specimens, AIEC strains were found in 21.7% of CD chronic lesions vs. in 6.2% of controls. In neoterminal ileal specimens, AIEC strains were found in 36.4% of CD early lesions (P = 0.034 vs. controls) and 22.2% of healthy mucosa of CD patients. In colonic specimens, AIEC strains were found in 3.7% of CD patients, 0% of UC patients, and 1.9% of controls. CONCLUSIONS: AIEC strains are associated specifically with ileal mucosa in CD.

Swidsinski A. · No affiliation provided · Inflamm Bowel Dis. · Pubmed #16917237 No free full text.

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