Probiotic Bacteria Regulate Intestinal Epithelial Permeability in Experimental Ileitis by a TNF-Dependent Mechanism
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{"title"=>"Probiotic bacteria regulate intestinal epithelial permeability in experimental ileitis by a TNF-dependent mechanism", "type"=>"journal", "authors"=>[{"first_name"=>"Daniele", "last_name"=>"Corridoni", "scopus_author_id"=>"55303810800"}, {"first_name"=>"Luca", "last_name"=>"Pastorelli", "scopus_author_id"=>"24484455700"}, {"first_name"=>"Benedetta", "last_name"=>"Mattioli", "scopus_author_id"=>"56554984500"}, {"first_name"=>"Silviu", "last_name"=>"Locovei", "scopus_author_id"=>"57199592484"}, {"first_name"=>"Dai", "last_name"=>"Ishikawa", "scopus_author_id"=>"55323105200"}, {"first_name"=>"Kristen O.", "last_name"=>"Arseneau", "scopus_author_id"=>"6602403733"}, {"first_name"=>"Marcello", "last_name"=>"Chieppa", "scopus_author_id"=>"9639838400"}, {"first_name"=>"Fabio", "last_name"=>"Cominelli", "scopus_author_id"=>"7006033187"}, {"first_name"=>"Theresa T.", "last_name"=>"Pizarro", "scopus_author_id"=>"6701374821"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "pmid"=>"22848704", "sgr"=>"84864336737", "pui"=>"365330233", "scopus"=>"2-s2.0-84864336737", "doi"=>"10.1371/journal.pone.0042067", "issn"=>"19326203"}, "id"=>"95045145-a447-3f44-b232-af565831bb31", "abstract"=>"BACKGROUND We previously showed that the probiotic mixture, VSL#3, prevents the onset of ileitis in SAMP/YitFc (SAMP) mice, and this effect was associated with stimulation of epithelial-derived TNF. The aim of this study was to determine the mechanism(s) of VSL#3-mediated protection on epithelial barrier function and to further investigate the \"paradoxical\" effects of TNF in preventing SAMP ileitis. METHODS Permeability was evaluated in SAMP mice prior to the onset of inflammation and during established disease by measuring transepithelial electrical resistance (TEER) on ex vivo-cultured ilea following exposure to VSL#3 conditioned media (CM), TNF or VSL#3-CM + anti-TNF. Tight junction (TJ) proteins were assessed by qRT-PCR, Western blot, and confocal microscopy, and TNFRI/TNFRII expression measured in freshly isolated intestinal epithelial cells (IEC) from SAMP and control AKR mice. RESULTS Culture with either VSL#3-CM or TNF resulted in decreased ileal paracellular permeability in pre-inflamed SAMP, but not SAMP with established disease, while addition of anti-TNF abrogated these effects. Modulation of the TJ proteins, claudin-2 and occludin, occurred with a significant decrease in claudin-2 and increase in occludin following stimulation with VSL#3-CM or TNF. TNF protein levels increased in supernatants of SAMP ilea incubated with VSL#3-CM compared to vehicle, while IEC-derived TNFR mRNA expression decreased in young, and was elevated in inflamed, SAMP versus AKR mice. CONCLUSIONS Our data demonstrate that the previously established efficacy of VSL#3 in preventing SAMP ileitis is due to direct innate and homeostatic effects of TNF on the gut epithelium, modulation of the TJ proteins, claudin-2 and occludin, and overall improvement of intestinal permeability.", "link"=>"http://www.mendeley.com/research/probiotic-bacteria-regulate-intestinal-epithelial-permeability-experimental-ileitis-tnfdependent-mec", "reader_count"=>68, "reader_count_by_academic_status"=>{"Unspecified"=>2, "Professor > Associate Professor"=>4, "Researcher"=>14, "Student > Doctoral Student"=>5, "Student > Ph. D. Student"=>11, "Student > Postgraduate"=>5, "Student > Master"=>8, "Other"=>4, "Student > Bachelor"=>10, "Lecturer"=>2, "Professor"=>3}, "reader_count_by_user_role"=>{"Unspecified"=>2, "Professor > Associate Professor"=>4, "Researcher"=>14, "Student > Doctoral Student"=>5, "Student > Ph. D. Student"=>11, "Student > Postgraduate"=>5, "Student > Master"=>8, "Other"=>4, "Student > Bachelor"=>10, "Lecturer"=>2, "Professor"=>3}, "reader_count_by_subject_area"=>{"Unspecified"=>5, "Biochemistry, Genetics and Molecular Biology"=>5, "Nursing and Health Professions"=>2, "Agricultural and Biological Sciences"=>31, "Medicine and Dentistry"=>18, "Arts and Humanities"=>1, "Chemistry"=>1, "Psychology"=>1, "Immunology and Microbiology"=>4}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>18}, "Chemistry"=>{"Chemistry"=>1}, "Psychology"=>{"Psychology"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>4}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>31}, "Nursing and Health Professions"=>{"Nursing and Health Professions"=>2}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>5}, "Unspecified"=>{"Unspecified"=>5}, "Arts and Humanities"=>{"Arts and Humanities"=>1}}, "reader_count_by_country"=>{"Belgium"=>1, "United States"=>2, "Italy"=>1, "United Kingdom"=>1}, "group_count"=>4}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/603406"], "description"=>"<p>(<i>A</i>), Representative confocal images (40X magnification) of ileal loops from young SAMP exposed to either VSL#3-CM, TNF or vehicle control, and immunofluorescent staining performed to detect occludin (upper panels) claudin-2 (lower panels). Epithelial occludin expression potently increased also in the villous tips after either VSL#3-CM or TNF stimulation in SAMP ilea. Claudin-2 primarily localized to the villous tips of SAMP ilea and decreased following VSL#3-CM, and particularly TNF, treatment. Results are representative of two independent experiments. (<i>B</i>), Representative Western blot of unfractionated IEC extracts isolated from SAMP mice treated with high dose VSL#3 showing increased expression of occludin and decreased claudin-2 protein levels compared to control. Results are representative of three independent experiments. (<i>C</i>), Representative Western blots demonstrating increased protein levels of occludin in membrane vs. cytosolic fractions of IEC from SAMP ileal loops treated with either VSL#3 or TNF compared to control. Claudin-2 protein was predominantly expressed in cytosolic vs. membrane fractions, and decreased following treatment with both VSL#3-CM and TNF compared to control. Results are representative of two independent experiments, pooling IEC from N = 3 mice/exp grp.</p>", "links"=>[], "tags"=>["epithelial", "claudin-2", "occludin"], "article_id"=>273888, "categories"=>["Chemistry", "Immunology"], "users"=>["Daniele Corridoni", "Luca Pastorelli", "Benedetta Mattioli", "Silviu Locovei", "Dai Ishikawa", "Kristen O. Arseneau", "Marcello Chieppa", "Fabio Cominelli", "Theresa T. Pizarro"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0042067.g004", "stats"=>{"downloads"=>0, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Localization_and_protein_expression_of_epithelial_claudin_2_and_occludin_following_in_vivo_treatment_with_VSL_3_CM_and_TNF_/273888", "title"=>"Localization and protein expression of epithelial claudin-2 and occludin following <i>in vivo</i> treatment with VSL#3-CM and TNF.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-07-25 01:04:48"}
  • {"files"=>["https://ndownloader.figshare.com/files/315129"], "description"=>"<div><h3>Background</h3><p>We previously showed that the probiotic mixture, VSL#3, prevents the onset of ileitis in SAMP/YitFc (SAMP) mice, and this effect was associated with stimulation of epithelial-derived TNF. The aim of this study was to determine the mechanism(s) of VSL#3-mediated protection on epithelial barrier function and to further investigate the “paradoxical” effects of TNF in preventing SAMP ileitis.</p> <h3>Methods</h3><p>Permeability was evaluated in SAMP mice prior to the onset of inflammation and during established disease by measuring transepithelial electrical resistance (TEER) on <em>ex vivo</em>-cultured ilea following exposure to VSL#3 conditioned media (CM), TNF or VSL#3-CM + anti-TNF. Tight junction (TJ) proteins were assessed by qRT-PCR, Western blot, and confocal microscopy, and TNFRI/TNFRII expression measured in freshly isolated intestinal epithelial cells (IEC) from SAMP and control AKR mice.</p> <h3>Results</h3><p>Culture with either VSL#3-CM or TNF resulted in decreased ileal paracellular permeability in pre-inflamed SAMP, but not SAMP with established disease, while addition of anti-TNF abrogated these effects. Modulation of the TJ proteins, claudin-2 and occludin, occurred with a significant decrease in claudin-2 and increase in occludin following stimulation with VSL#3-CM or TNF. TNF protein levels increased in supernatants of SAMP ilea incubated with VSL#3-CM compared to vehicle, while IEC-derived TNFR mRNA expression decreased in young, and was elevated in inflamed, SAMP versus AKR mice.</p> <h3>Conclusions</h3><p>Our data demonstrate that the previously established efficacy of VSL#3 in preventing SAMP ileitis is due to direct innate and homeostatic effects of TNF on the gut epithelium, modulation of the TJ proteins, claudin-2 and occludin, and overall improvement of intestinal permeability.</p> </div>", "links"=>[], "tags"=>["probiotic", "intestinal", "epithelial", "permeability", "ileitis", "tnf-dependent"], "article_id"=>122183, "categories"=>["Chemistry", "Immunology"], "users"=>["Daniele Corridoni", "Luca Pastorelli", "Benedetta Mattioli", "Silviu Locovei", "Dai Ishikawa", "Kristen O. Arseneau", "Marcello Chieppa", "Fabio Cominelli", "Theresa T. Pizarro"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0042067", "stats"=>{"downloads"=>1, "page_views"=>49, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Probiotic_Bacteria_Regulate_Intestinal_Epithelial_Permeability_in_Experimental_Ileitis_by_a_TNF_Dependent_Mechanism/122183", "title"=>"Probiotic Bacteria Regulate Intestinal Epithelial Permeability in Experimental Ileitis by a TNF-Dependent Mechanism", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-07-25 00:36:23"}
  • {"files"=>["https://ndownloader.figshare.com/files/603238"], "description"=>"<p>Supernatants were collected from cultured ilea that were exposed apically to VSL#3-CM and cytokine protein levels measured as described in Methods and Materials. *<i>p</i><0.05 and **<i>p</i><0.01 vs. vehicle control or age- and treatment-matched AKR. Results are representative of two independent experiments (N = 6/exp grp) and expressed as mean ± SEM.</p>", "links"=>[], "tags"=>["ileal", "innate-type", "cytokine", "secretion"], "article_id"=>273716, "categories"=>["Chemistry", "Immunology"], "users"=>["Daniele Corridoni", "Luca Pastorelli", "Benedetta Mattioli", "Silviu Locovei", "Dai Ishikawa", "Kristen O. Arseneau", "Marcello Chieppa", "Fabio Cominelli", "Theresa T. Pizarro"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0042067.g002", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Increased_ileal_innate_type_cytokine_secretion_following_VSL3_treatment_/273716", "title"=>"Increased ileal innate-type cytokine secretion following VSL3# treatment.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-07-25 01:01:56"}
  • {"files"=>["https://ndownloader.figshare.com/files/603548"], "description"=>"<p>(<i>A</i>), TNFRI and II mRNA expression measured by qRT-PCR was decreased in freshly isolated IEC from 4-wk-old SAMP prior to the onset of inflammation, but increased in >20-wk-old SAMP with established disease compared to age-matched AKR control mice (N = 6/exp grp). *<i>p</i><0.05 and **<i>p</i><0.01. (<i>B</i>), Epithelial paracellular permeability was assessed by measuring ΔTEER on <i>ex vivo</i> cultured full-thickness ilea from mice deficient in both TNFRI and TNFRII (TNFR KO) and WT controls after 1 h exposure to VSL#3-CM or TNF (N = 6/exp grp). *<i>p</i><0.05 vs. WT vehicle cont.</p>", "links"=>[], "tags"=>["ilea", "differentially"], "article_id"=>274031, "categories"=>["Chemistry", "Immunology"], "users"=>["Daniele Corridoni", "Luca Pastorelli", "Benedetta Mattioli", "Silviu Locovei", "Dai Ishikawa", "Kristen O. Arseneau", "Marcello Chieppa", "Fabio Cominelli", "Theresa T. Pizarro"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0042067.g005", "stats"=>{"downloads"=>0, "page_views"=>19, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_SAMP_ilea_differentially_express_TNFRs_which_are_essential_to_the_effects_of_VSL_3_CM_and_TNF_/274031", "title"=>"SAMP ilea differentially express TNFRs, which are essential to the effects of VSL#3-CM and TNF.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-07-25 01:07:11"}
  • {"files"=>["https://ndownloader.figshare.com/files/603149"], "description"=>"<p>Epithelial paracellular permeability was assessed by measuring TEER on <i>ex vivo</i> cultured full-thickness ilea from (<i>A</i>), 3- to 4-wk old SAMP, prior to the onset of inflammation, (<i>B</i>), older (>20 wks) SAMP with established ileitis, and age-matched AKR controls at T<sub>0</sub> and after 1 h (ΔTEER) exposure to VSL#3-CM or TNF (N = 6/exp grp). **<i>p</i><0.01 vs. age- and treatment-matched AKR, *<i>p</i><0.05 vs. vehicle cont. (<i>C</i>), Addition of a neutralizing antibody against TNF abrogated the positive effects of VSL#3 on ΔTEER in pre-inflamed ilea from young SAMP (N = 7 exp grp). *<i>p</i><0.05 vs. vehicle cont. Results are combined from two independent experiments and expressed as mean ± SEM.</p>", "links"=>[], "tags"=>["decreases", "epithelial", "paracellular", "permeability", "tnf-dependent", "ilea", "pre-inflamed", "samp"], "article_id"=>273611, "categories"=>["Chemistry", "Immunology"], "users"=>["Daniele Corridoni", "Luca Pastorelli", "Benedetta Mattioli", "Silviu Locovei", "Dai Ishikawa", "Kristen O. Arseneau", "Marcello Chieppa", "Fabio Cominelli", "Theresa T. Pizarro"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0042067.g001", "stats"=>{"downloads"=>0, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_VSL_3_decreases_epithelial_paracellular_permeability_in_a_TNF_dependent_manner_on_ilea_from_pre_inflamed_SAMP_mice_/273611", "title"=>"VSL#3 decreases epithelial paracellular permeability in a TNF-dependent manner on ilea from pre-inflamed SAMP mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-07-25 01:00:11"}
  • {"files"=>["https://ndownloader.figshare.com/files/603299"], "description"=>"<p>mRNA transcript levels of TJ proteins (claudins 1–4, ZO-1, and occludin) were measured by qRT-PCR and expressed relative to ß-actin in full-thickness ilea from experimental mice following <i>ex vivo</i> culture with either VSL#3-CM or TNF. The greatest differences in expression were seen in claudin-2 (decrease) and occludin (increase) after either VSL#3-CM or TNF stimulation, and in claudin-3 (decrease) after TNF stimulation only. No differences in expression were noted for claudins 1, 4, and ZO-1 following exposure to either VSL#3-CM or TNF. *<i>p</i><0.05 vs. vehicle cont. Results are representative of two independent experiments (N = 6/exp group) and expressed as mean ± SEM.</p>", "links"=>[], "tags"=>["tnf", "modulate", "tj", "ilea", "pre-inflamed", "samp"], "article_id"=>273779, "categories"=>["Chemistry", "Immunology"], "users"=>["Daniele Corridoni", "Luca Pastorelli", "Benedetta Mattioli", "Silviu Locovei", "Dai Ishikawa", "Kristen O. Arseneau", "Marcello Chieppa", "Fabio Cominelli", "Theresa T. Pizarro"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0042067.g003", "stats"=>{"downloads"=>1, "page_views"=>20, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_VSL_3_and_TNF_modulate_TJ_protein_expression_in_ilea_from_pre_inflamed_SAMP_mice_/273779", "title"=>"VSL#3 and TNF modulate TJ protein expression in ilea from pre-inflamed SAMP mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-07-25 01:02:59"}

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Relative Metric

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