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  • 1.
    Rangel, Ignacio
    et al.
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Sundin, Johanna
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Fuentes, S.
    Laboratory of Microbiology, Wageningen University, Wageningen, The Netherlands.
    Repsilber, Dirk
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    de Vos, W. M.
    Laboratory of Microbiology, Wageningen University, Wageningen, The Netherlands; Departments of Bacteriology & Immunology and Veterinary Biosciences, University of Helsinki, Helsinki, Finland .
    Brummer, Robert Jan
    Örebro University, School of Medicine, Örebro University, Sweden.
    The relationship between faecal-associated and mucosal-associated microbiota in irritable bowel syndrome patients and healthy subjects2015In: Alimentary Pharmacology and Therapeutics, ISSN 0269-2813, E-ISSN 1365-2036, Vol. 42, no 10, p. 1211-1221Article in journal (Refereed)
    Abstract [en]

    Background: The faecal-associated microbiota is commonly seen as a surrogate of the mucosal-associated microbiota. However, previous studies indicate that they are different. Furthermore, analyses of the mucosal microbiota are commonly done after standard bowel cleansing, affecting the microbial composition.

    Aim: To compare the mucosal-associated microbiota, obtained from unprepared colon, with faecal-associated microbiota in healthy subjects and irritable bowel syndrome (IBS) patients.

    Methods: Faecal and mucosal biopsies were obtained from 33 IBS patients and 16 healthy controls. Of IBS patients, 49% belonged to the diarrhoea-predominant subgroup and 80% suffered from IBS symptoms during at least 5 years. Biopsies were collected from unprepared sigmoid colon and faecal samples a day before colonoscopy. Microbiota analyses were performed with a phylogenetic microarray and redundancy discriminant analysis.

    Results: The composition of the mucosal- and the faecal-associated microbiota in unprepared sigmoid colon differs significantly (P = 0.002). Clinical characteristics of IBS did not correlate with this difference. Bacteroidetes dominate the mucosal-associated microbiota. Firmicutes, Actinobacteria and Proteobacteria dominate the faecal-associated microbiota. Healthy subjects had a significantly higher (P < 0.005) abundance (1.9%) of the bacterial group uncultured Clostridiales I in the mucosal-associated microbiota than IBS patients (0.3%). Bacterial diversity was higher in faecal- compared with mucosal-associated microbiota in IBS patients (P < 0.005). No differences were found in healthy subjects.

    Conclusions: Differences in the mucosal-associated microbiota between healthy individuals and IBS patients are minimal (one bacterial group) compared to differences in the faecal microbiota of both groups (53 bacterial groups). Microbial aberrations characterising IBS are more pronounced in the faeces than in the mucosa.

  • 2.
    Rangel, Ignacio
    et al.
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Sundin, Johanna
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Fuentes, Susana
    Wageningen University, Wageningen, The Netherlands.
    Repsilber, Dirk
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    de Vos, Willem M.
    Wageningen University, Wageningen, The Netherlands; University of Helsinki, Finland.
    Brummer, Robert J.
    Örebro University, School of Medicine, Örebro University, Sweden.
    Mucosal-associated microbiota differs less than fecal-associated microbiota between Irritable Bowel Syndrome patients and healthy subjectsManuscript (preprint) (Other academic)
  • 3.
    Sundin, Johanna
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Microbe-host interactions in post-infectious irritable bowel syndrome2015Doctoral thesis, comprehensive summary (Other academic)
    List of papers
    1. Aberrant mucosal lymphocyte number and subsets in the colon of post-infectious irritable bowel syndrome patients
    Open this publication in new window or tab >>Aberrant mucosal lymphocyte number and subsets in the colon of post-infectious irritable bowel syndrome patients
    Show others...
    2014 (English)In: Scandinavian Journal of Gastroenterology, ISSN 0036-5521, E-ISSN 1502-7708, Vol. 49, no 9, p. 1068-1075Article in journal (Refereed) Published
    Abstract [en]

    Background: Irritable bowel syndrome (IBS) is characterized by chronic abdominal symptoms such as pain, discomfort, and altered bowel habits. A subset of IBS patients, denoted as post-infectious IBS (PI-IBS) patients, develop symptoms after an enteric infection. Distinct abnormalities in the gut mucosa, including mucosal inflammation, have been proposed to contribute to or be the cause of PI-IBS. This study investigated lymphocyte subsets in PI-IBS patients compared to healthy controls.

    Materials and methods: Ten PI-IBS patients and nine healthy controls participated. All PI-IBS patients met the Rome III diagnostic criteria for IBS and reported sustained symptoms at least 1 year after an episode of acute gastroenteritis. Intraepithelial lymphocytes and lamina propria lymphocytes (LPLs), isolated from mucosal tissue samples, were stained and analyzed for a comprehensive set of cell markers using flow cytometry.

    Results: The number of LPLs in PI-IBS was significantly increased compared to those in healthy controls (p < 0.05). PI-IBS patients showed significantly increased proportions of CD45RO(+) CD4(+) activated/memory T cells (p < 0.05) and double-positive CD4(+) CD8(+) cells (p < 0.05), respectively, in the lamina propria. The number of CD19(+) LPLs was decreased in PI-IBS patients compared to healthy controls (p < 0.001).

    Conclusion: This study presents new evidence that PI-IBS is associated with a sustained aberrant mucosal immune response and support future studies of anti-inflammatory or immune-modulating treatments in these patients.

    Place, publisher, year, edition, pages
    Informa Healthcare, 2014
    Keywords
    cell biology, gastrointestinal, infections, health economy, immunology
    National Category
    Gastroenterology and Hepatology
    Identifiers
    urn:nbn:se:oru:diva-35383 (URN)10.3109/00365521.2014.926982 (DOI)000340829900006 ()24919810 (PubMedID)2-s2.0-84906318425 (Scopus ID)
    Note

    Funding Agency:

    Medical Faculty, Örebro University

    Available from: 2014-06-17 Created: 2014-06-17 Last updated: 2019-03-26Bibliographically approved
    2. Altered faecal and mucosal microbial composition in post-infectious irritable bowel syndrome patients correlates with mucosal lymphocyte phenotypes and psychological distress
    Open this publication in new window or tab >>Altered faecal and mucosal microbial composition in post-infectious irritable bowel syndrome patients correlates with mucosal lymphocyte phenotypes and psychological distress
    Show others...
    2015 (English)In: Alimentary Pharmacology and Therapeutics, ISSN 0269-2813, E-ISSN 1365-2036, Vol. 41, no 4, p. 342-351Article in journal (Refereed) Published
    Abstract [en]

    Background: A subset of irritable bowel syndrome (IBS) patients, denoted post-infectious IBS (PI-IBS), develop symptoms after an enteric infection. Bacterial dysbiosis and mucosal inflammation have been proposed to be involved in the pathophysiology of this entity.

    Aim: To characterise the mucosal and faecal microbiota in PI-IBS, general IBS and healthy controls, and to investigate associations between the microbiota and the mucosal immune system.

    Methods: Mucosal biopsies and faeces were collected from 13 PI-IBS patients, 19 general IBS patients and 16 healthy controls. Global bacterial composition was determined by generating 16S rRNA amplicons that were examined by phylogenetic microarray hybridisation, principal component and redundancy analysis. We correlated previously reported lymphocyte proportions with the microbiota.

    Results: Faecal microbiota composition of PI-IBS patients differed significantly from both general IBS patients and healthy controls (P < 0.02). Both mucosal (P < 0.01) and faecal (P = 0.05) microbial diversity were reduced in PI-IBS compared to healthy controls. In the intraepithelial lymphocytes the previously published proportion of CD8+ CD45RA+ was negatively correlated with mucosal microbial diversity (P < 0.005). The previously published number of lamina propria lymphocytes was negatively correlated with mucosal microbial diversity (P < 0.05). Faecal microbial diversity was significantly negatively correlated with the Hospital Anxiety and Depression scale (P < 0.05).

    Conclusions: We present data that distinguishes the intestinal microbiota of PI-IBS patients from that of both general IBS patients and HC. The microbial composition is significantly associated with the HADs score and alterations in lymphocyte subsets proportions.

    Place, publisher, year, edition, pages
    Hoboken, USA: Wiley-Blackwell, 2015
    National Category
    Cell and Molecular Biology Gastroenterology and Hepatology
    Identifiers
    urn:nbn:se:oru:diva-39955 (URN)10.1111/apt.13055 (DOI)000348731200002 ()25521822 (PubMedID)2-s2.0-84921438829 (Scopus ID)
    Note

    Funding Agencies:

    Medical Faculty, Orebro University

    Spinoza award of the Netherlands Organization for Scientific Research

    Gravity grant (SIAM) of the Netherlands Organization for Scientific Research

    Available from: 2014-12-22 Created: 2014-12-22 Last updated: 2018-01-11Bibliographically approved
    3. Mucosal-associated microbiota differs less than fecal-associated microbiota between Irritable Bowel Syndrome patients and healthy subjects
    Open this publication in new window or tab >>Mucosal-associated microbiota differs less than fecal-associated microbiota between Irritable Bowel Syndrome patients and healthy subjects
    Show others...
    (English)Manuscript (preprint) (Other academic)
    National Category
    Gastroenterology and Hepatology
    Identifiers
    urn:nbn:se:oru:diva-43135 (URN)
    Available from: 2015-03-02 Created: 2015-03-02 Last updated: 2017-10-17Bibliographically approved
    4. Cytokine response after stimulation with key commensal bacteria differ in post-­infectious irritable bowel syndrome (PI-­IBS) patients compared to healthy subjects
    Open this publication in new window or tab >>Cytokine response after stimulation with key commensal bacteria differ in post-­infectious irritable bowel syndrome (PI-­IBS) patients compared to healthy subjects
    (English)Manuscript (preprint) (Other academic)
    National Category
    Gastroenterology and Hepatology
    Identifiers
    urn:nbn:se:oru:diva-43133 (URN)
    Available from: 2015-03-02 Created: 2015-03-02 Last updated: 2017-10-17Bibliographically approved
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  • 4.
    Sundin, Johanna
    et al.
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Brummer, Robert
    Örebro University, School of Medicine, Örebro University, Sweden.
    Hultgren Hörnquist, Elisabet
    Örebro University, School of Medicine, Örebro University, Sweden.
    Rangel, Ignacio
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Increased number of double positive CD3+ CD8+ CD4+ lamina propria T lymphocyte in gut mucosa of post infectious IBS patients compared to healthy controls2012In: Neurogastroenterology and Motility, ISSN 1350-1925, E-ISSN 1365-2982, Vol. 24, no Suppl. 2, p. 104-105Article in journal (Other academic)
    Abstract [en]

    Objective: Irritable bowel syndrome (IBS) developed after a gastroenteritis is denoted post infectious IBS (PI-IBS) and is thought to represent a specific patho-physiological entity. Naive CD8+CD45RA+cytotoxic T lymphocytes recognize antigen derived from intracellular bacteria and viruses. Naive CD4+CD45RA+helper T lymphocytes are activated by the antigen from extracellular microorganisms with the help of antigen-presenting cells. Activated CD4+CD45RO+helper T lymphocytes help phagocytes to kill microbes and activate naive B lymphocytes.

    Aim: To characterize subsets of mucosal lymphocytes in PI-IBS with flow cytometry analysis as a lead in identifying new therapeutic methods.

    Methods: 5 PI-IBS patients and 6 healthy individuals were recruited. We performed a distal colonoscopy without bowel cleansing or other preparation. Lamina propria lymphocytes (LPL) and intra-epithelial lymphocytes (IEL) were isolated from sigmodal biopsies and sub classified by CD3, CD4, CD8, CD45RO and CD45RA with flow cytometry.

    Results: We observed a significant deference (P < 0.01, two-tailed Mann–Whitney test) in double positive CD3+CD8+CD4+LPL between PI-IBS patient and healthy controls. We also observed a trend towards increased frequency of CD3+CD4+LPL in PI-IBS patients. Additionally, PI-IBS patients showed an increased frequency of CD3+CD8+LPL and IEL. The proportion of memory / activated CD45RO+CD4+LPL was higher in PI-IBS patients, and its proportion of the CD8+population was lower compared with the healthy controls. On the contrary the proportion of naive CD45RA+CD4+LPL was lower in PI-IBS patients compared with healthy controls. However, we found no differences in the distribution naı¨ve / activated CD4+ and CD8+IEL between PI-IBS patients and healthy controls.

    Conclusion: The difference in double positive (DP) CD3+CD8+CD4+LPL seen between PI-IBS patient and healthy controls is consistent with findings of increased number DP T cells in target organs in immuno-inflammatory conditions. Our analysis revealed that the normal gut has a greater variation in the prevalence of CD3+CD4+and CD4+CD45RO+LPL than that of PI-IBS patients. These findings confirm aberrant mucosal subsets of lymphocytes. However, more subjects need to be included in the study in order to draw firm conclusions.

  • 5. Sundin, Johanna
    et al.
    Kumawat, Ashok Kumar
    Rangel, I.
    Brummer, Robert
    Hultgren Hörnquist, Elisabet
    Örebro University, School of Medicine, Örebro University, Sweden.
    Karakterisering av T-lymfocyter från tarmmukosan hos patienter med postinfektiös IBS2012Conference paper (Other academic)
  • 6.
    Sundin, Johanna
    et al.
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Rangel, Ignacio
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Fuentes, S
    Department of Microbiology, Wageningen University, Wageningen,The Netherlands.
    Jong, Heikamp-de
    Department of Microbiology, Wageningen University, Wageningen,The Netherlands.
    Hultgren-Hörnquist, Elisabeth
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    de Vos, W M
    Department of Microbiology, Wageningen University, Wageningen,The Netherlands; Departments of Bacterology & Immunology and Veterinary Biosciences, University of Helsinki, Helsinki, Finland.
    Brummer, Robert-Jan
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Altered faecal and mucosal microbial composition in post-infectious irritable bowel syndrome patients correlates with mucosal lymphocyte phenotypes and psychological distress2015In: Alimentary Pharmacology and Therapeutics, ISSN 0269-2813, E-ISSN 1365-2036, Vol. 41, no 4, p. 342-351Article in journal (Refereed)
    Abstract [en]

    Background: A subset of irritable bowel syndrome (IBS) patients, denoted post-infectious IBS (PI-IBS), develop symptoms after an enteric infection. Bacterial dysbiosis and mucosal inflammation have been proposed to be involved in the pathophysiology of this entity.

    Aim: To characterise the mucosal and faecal microbiota in PI-IBS, general IBS and healthy controls, and to investigate associations between the microbiota and the mucosal immune system.

    Methods: Mucosal biopsies and faeces were collected from 13 PI-IBS patients, 19 general IBS patients and 16 healthy controls. Global bacterial composition was determined by generating 16S rRNA amplicons that were examined by phylogenetic microarray hybridisation, principal component and redundancy analysis. We correlated previously reported lymphocyte proportions with the microbiota.

    Results: Faecal microbiota composition of PI-IBS patients differed significantly from both general IBS patients and healthy controls (P < 0.02). Both mucosal (P < 0.01) and faecal (P = 0.05) microbial diversity were reduced in PI-IBS compared to healthy controls. In the intraepithelial lymphocytes the previously published proportion of CD8+ CD45RA+ was negatively correlated with mucosal microbial diversity (P < 0.005). The previously published number of lamina propria lymphocytes was negatively correlated with mucosal microbial diversity (P < 0.05). Faecal microbial diversity was significantly negatively correlated with the Hospital Anxiety and Depression scale (P < 0.05).

    Conclusions: We present data that distinguishes the intestinal microbiota of PI-IBS patients from that of both general IBS patients and HC. The microbial composition is significantly associated with the HADs score and alterations in lymphocyte subsets proportions.

  • 7.
    Sundin, Johanna
    et al.
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Rangel, Ignacio
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden. Örebro University, School of Medicine, Örebro University, Sweden.
    Kumawat, Ashok K
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Hultgren-Hörnquist, Elisabeth
    Örebro University, School of Medicine, Örebro University, Sweden.
    Brummer, Robert J
    Örebro University, School of Medicine, Örebro University, Sweden. Örebro University Hospital.
    Aberrant mucosal lymphocyte number and subsets in the colon of post-infectious irritable bowel syndrome patients2014In: Scandinavian Journal of Gastroenterology, ISSN 0036-5521, E-ISSN 1502-7708, Vol. 49, no 9, p. 1068-1075Article in journal (Refereed)
    Abstract [en]

    Background: Irritable bowel syndrome (IBS) is characterized by chronic abdominal symptoms such as pain, discomfort, and altered bowel habits. A subset of IBS patients, denoted as post-infectious IBS (PI-IBS) patients, develop symptoms after an enteric infection. Distinct abnormalities in the gut mucosa, including mucosal inflammation, have been proposed to contribute to or be the cause of PI-IBS. This study investigated lymphocyte subsets in PI-IBS patients compared to healthy controls.

    Materials and methods: Ten PI-IBS patients and nine healthy controls participated. All PI-IBS patients met the Rome III diagnostic criteria for IBS and reported sustained symptoms at least 1 year after an episode of acute gastroenteritis. Intraepithelial lymphocytes and lamina propria lymphocytes (LPLs), isolated from mucosal tissue samples, were stained and analyzed for a comprehensive set of cell markers using flow cytometry.

    Results: The number of LPLs in PI-IBS was significantly increased compared to those in healthy controls (p < 0.05). PI-IBS patients showed significantly increased proportions of CD45RO(+) CD4(+) activated/memory T cells (p < 0.05) and double-positive CD4(+) CD8(+) cells (p < 0.05), respectively, in the lamina propria. The number of CD19(+) LPLs was decreased in PI-IBS patients compared to healthy controls (p < 0.001).

    Conclusion: This study presents new evidence that PI-IBS is associated with a sustained aberrant mucosal immune response and support future studies of anti-inflammatory or immune-modulating treatments in these patients.

  • 8.
    Sundin, Johanna
    et al.
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Rangel, Ignacio
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Repsilber, Dirk
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Brummer, Robert J.
    Örebro University, School of Medicine, Örebro University, Sweden.
    Cytokine response after stimulation with key commensal bacteria differ in post-­infectious irritable bowel syndrome (PI-­IBS) patients compared to healthy subjectsManuscript (preprint) (Other academic)
  • 9.
    Sundin, Johanna
    et al.
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Rangel, Ignacio
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Repsilber, Dirk
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Brummer, Robert-Jan
    Örebro University, School of Medicine, Örebro University, Sweden.
    Cytokine Response after Stimulation with Key Commensal Bacteria Differ in Post-Infectious Irritable Bowel Syndrome (PI-IBS) Patients Compared to Healthy Controls2015In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 10, no 9, article id e0134836Article in journal (Refereed)
    Abstract [en]

    Background: Microbial dysbiosis and prolonged immune activation resulting in low-grade inflammation and intestinal barrier dysfunction have been suggested to be underlying causes of post-infectious irritable bowel syndrome (PI-IBS). The aim of this study was to evaluate the difference in cytokine response between mucosal specimens of PI-IBS patients and healthy controls (HC) after ex vivo stimulation with key anaerobic bacteria.

    Methods: Colonic biopsies from 11 PI-IBS patients and 10 HC were stimulated ex vivo with the commensal bacteria Bacteroides ovatus, Ruminococcus gnavus, Akkermansia muciniphila, Subdoligranulum variabile and Eubacterium limosum, respectively. The cytokine release (IL-1 beta, IL-2, IL-8, IL-10, IL-13, IL-17, TNF-alpha and IFN-gamma) in stimulation supernatants was analyzed using the LUMINEX assay. Comparison of cytokine release between PI-IBS patients and healthy controls was performed taking both unstimulated and bacterially stimulated mucosal specimens into account.

    Key Results: IL-13 release from mucosal specimens without bacterial stimulation was significantly lower in PI-IBS patients compared to HC (p < 0.05). After stimulation with Subdoligranulum variabile, IL-1 beta release from PI-IBS patients was significantly increased compared to HC (p < 0.05). Stimulation with Eubacterium limosum resulted in a significantly decreased IL-10 release in HC compared to PI-IBS patients (p < 0.05) and a tendency to decreased IL-13 release in HC compared to PI-IBS patients (p = 0.07).

    Conclusions & Inferences: PI-IBS patients differ from HC with regard to cytokine release ex vivo after stimulation with selected commensal bacteria. Hence, our results support that the pathogenesis of PI-IBS comprises an altered immune response against commensal gut microbes.

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