Mucosa Of IBD Patients Research Articles

Influence of vitamin D on the expression of mRNA of cytokines in the mucosa of inflammatory bowel disease patients.

The aim of this study was to analyze the influence of 25(OH)VD serum concentration on the expression of mRNA cytokines (IL-6, IL-8, IL-12, IL-17, IL-23, TNFα, CCR1, CCR2, CCR5, CCR9, CCL5, TLR2, TLR4, TLR5, CD207 ,CD206, FoxP3) in mucosa of IBD patients. The cohort consisted of 86 IBD patients (48 CD and 38UC) followed at the IBD center of University Hospital Bratislava-Ruzinov. We performed colonoscopy in each patient and took biopsies from mucosa of sigma and terminal ileum. Serum concentration of 25(OH)VD was assessed at the time of colonoscopy. mRNA was extracted from mucosal biopsy samples for each cytokine. Then we analyzed the correlation between VD and the expression of mRNA of cytokines from biopsies samples. In CD we observed asignificant positive correlation of serum concentration 25(OH)VD and the expression mRNA level of IL-6. There was also trend towards significant positive correlation of the expression mRNA of TNFα, IL-10, IL-23, TLR 2 in inflamed mucosa of terminal ileum as well as the expression mRNA of CCR5 and CCR1 in non-inflamed mucosa from terminal ileum. We also found atrend towards positive correlation between 25(OH)VD and the expression mRNA of IL-23, TLR4, CD 207, CCR1, CCR5 and CD 206 in non-inflamed mucosa of sigma in UC.VD significantly correlated with the levels of expression of several inflammatory cytokines including TNFα in colonic mucosa of patients with IBD (Tab. 4, Fig. 3, Ref. 31).

P074 Expression of CCL20 and its corresponding receptor CCR6 is located to epithelium and infiltrating cells in the mucosa of IBD patients

presence of myofibroblasts in all layers of the intestinal wall. Ulcerative colitis (UC) is classically considered to be a purely mucosal disease although rarely transmural complications such as strictures and stenosis develop. Fibrogenesis in UC has not been studied systematically yet and may be a neglected phenomenon. We therefore investigated whether there is a different fibrotic load in acute vs longstanding UC and whether the degree of fibrosis in UC correlates with the severity of inflammation. Methods: Colectomy specimens from all UC patients operated at the AMC between 2007 2012 were reviewed. Specimens from patients with recent onset refractory therapy UC (diagnosis 10 years) were selected. Patients operated for colon cancer without UC served as controls. On HE 29% vs 33% vs 54%, p = 0.009). Between acute and late UC there was no difference in collagen deposition, however between UC together and controls there was more collagen I expression in the mucosa, MM and muscularis externa (50% vs 10%, p = 0.02; 28% vs 10%, p = 0.048; 71% vs 20%, p = 0.003). In both early and long UC duration, we did not find a correlation between inflammation or collagen deposition or MM thickness. There was a negative correlation between inflammation and aSMA positivity in the submucosa (R = 0.51, p = 0.003) and MM (R = 0.37, p = 0.04). Conclusions: In our series of colectomy specimens, there is more collagen deposition in UC than in controls. No association between disease duration and increased collagen deposition could be found. Expression of aSMA however is lower in UC and correlates with inflammation. Fibrosis in UC does not appear to increase significantly over time.

CS15-7. Decrease of IL-17 and Gm-Csf in Intestinal Mucosa of Ibd Patients is Associated With Reduction of CD68+ Macrophages and Clinical Remission in Response to Anti-TNF Therapy

CS15-7. Decrease of IL-17 and Gm-Csf in Intestinal Mucosa of Ibd Patients is Associated With Reduction of CD68+ Macrophages and Clinical Remission in Response to Anti-TNF Therapy

IL23R, ATG16L1, IRGM, OCTN1, and OCTN2 mRNA expression in inflamed and noninflamed mucosa of IBD patients

IL23R, ATG16L1, IRGM, OCTN1, and OCTN2 mRNA expression in inflamed and noninflamed mucosa of IBD patients

W1216 IL23R, ATG16L1, IRGM, and Octns mRNA Expression in Inflamed and Non-Inflamed Mucosa of IBD Patients

W1216 IL23R, ATG16L1, IRGM, and Octns mRNA Expression in Inflamed and Non-Inflamed Mucosa of IBD Patients

T1686 Carrageenan-Mediated Increases in NfκB Involve the Ikkα, β, and γ Components of the Ikk Signalosome

INTRODUCTION: An involvement of the intestinal lymphatic systemhas been long suggested in IBD(1). New markers have become available to distinguish lymphatic vessels from blood vessels. AIMS & METHODS: The aim of the study was to examine the distribution of lymphatic vessels within macroscopically inflamed and non-inflamed colonic mucosa of IBD patients (18 CD, 17 ulcerative colitis (UC)) and compare it with healthy controls (9) using immunohistochemistry and real-time PCR for podoplanin. Quantification of lymphatic vessels was performed on podoplanin stained sections using Weibel graticule in 7 fields randomly selected in all patients. Lymphatic vessel density was expressed as mean ±SEM surface area (SA). Levels of podoplanin mRNA were expressed as relative mean ±SEM unit. For statistical analysis, a permutation test was used. RESULTS: In CD patients, the number of lymphatics was significantly increased in non-inflamed (SA 1.89%±0.28, p <0.01) and inflamed colonic mucosa (SA 3.0%±0.30, p<0.001) compared to controls (SA 0.36%±0.14). Podoplanin mRNA levels were similar in non inflamed areas (13.75±1.97, NS) compared to controls (11.98±3.13) whereas a six fold increase was seen in inflamed areas (74.70±29.80, p<0.05). In UC patients, the number of lymphatics was significantly increased in non-inflamed (SA 2.38%±0.27, p<0.001) and inflamed colonic mucosa (SA 2.77%±0.28, p<0.001) compared to controls (SA 0.36%±0.14). An increase in podoplanin mRNA levels was seen both in non inflamed (17.42±6.72, NS) and inflamed areas (117.43±39, p<0.01) compared to controls (11.98±3.13). In control patients, lymphatics were only present close to or under the muscularis mucosae contrasting with a diffuse presence throughout the mucosa in both CD and UC patients. CONCLUSION: A strong lymphangiogenesis occurs within the colonic mucosa of IBD patients together with a change in distribution and appearance of lymphatics in the upper half of the lamina propria. The present data further suggest that lymphangiogenesis play a role in the pathogenesis of IBD. Furthermore, this distribution challenges the established dogma stating the lack of lymphatic vessels in colonic mucosa. (1)Van Kruiningen HJ, Colombel JF. The forgotten role of lymphangitis in Crohn's disease. Gut 2008;57:1-4.

T1685 Mucosal Lymphangiogenesis: An Integral Component of Inflammatory Bowel Disease Pathogenesis

INTRODUCTION: An involvement of the intestinal lymphatic systemhas been long suggested in IBD(1). New markers have become available to distinguish lymphatic vessels from blood vessels. AIMS & METHODS: The aim of the study was to examine the distribution of lymphatic vessels within macroscopically inflamed and non-inflamed colonic mucosa of IBD patients (18 CD, 17 ulcerative colitis (UC)) and compare it with healthy controls (9) using immunohistochemistry and real-time PCR for podoplanin. Quantification of lymphatic vessels was performed on podoplanin stained sections using Weibel graticule in 7 fields randomly selected in all patients. Lymphatic vessel density was expressed as mean ±SEM surface area (SA). Levels of podoplanin mRNA were expressed as relative mean ±SEM unit. For statistical analysis, a permutation test was used. RESULTS: In CD patients, the number of lymphatics was significantly increased in non-inflamed (SA 1.89%±0.28, p <0.01) and inflamed colonic mucosa (SA 3.0%±0.30, p<0.001) compared to controls (SA 0.36%±0.14). Podoplanin mRNA levels were similar in non inflamed areas (13.75±1.97, NS) compared to controls (11.98±3.13) whereas a six fold increase was seen in inflamed areas (74.70±29.80, p<0.05). In UC patients, the number of lymphatics was significantly increased in non-inflamed (SA 2.38%±0.27, p<0.001) and inflamed colonic mucosa (SA 2.77%±0.28, p<0.001) compared to controls (SA 0.36%±0.14). An increase in podoplanin mRNA levels was seen both in non inflamed (17.42±6.72, NS) and inflamed areas (117.43±39, p<0.01) compared to controls (11.98±3.13). In control patients, lymphatics were only present close to or under the muscularis mucosae contrasting with a diffuse presence throughout the mucosa in both CD and UC patients. CONCLUSION: A strong lymphangiogenesis occurs within the colonic mucosa of IBD patients together with a change in distribution and appearance of lymphatics in the upper half of the lamina propria. The present data further suggest that lymphangiogenesis play a role in the pathogenesis of IBD. Furthermore, this distribution challenges the established dogma stating the lack of lymphatic vessels in colonic mucosa. (1)Van Kruiningen HJ, Colombel JF. The forgotten role of lymphangitis in Crohn's disease. Gut 2008;57:1-4.

S1746 IL-10/TNFα Antagonism in the Intestinal Mucosa of IBD Patients. Relation with Steroid Response

S1746 IL-10/TNFα Antagonism in the Intestinal Mucosa of IBD Patients. Relation with Steroid Response

Evaluation of the MLH1 I219V alteration in DNA mismatch repair activity and ulcerative colitis

Inflammatory bowel diseases (IBDs; ulcerative colitis, UC, and Crohn's disease, CD) show familial clustering suggestive of a genetic background. A linkage susceptibility region for these diseases (IBD9) lies on chromosome 3p and includes the DNA mismatch repair gene MLH1. Loss of MLH1 confers the characteristic microsatellite instability (MSI) phenotype which is also frequently found in the mucosa of IBD patients. A common germline alteration of MLH1 (655A>G) results in the amino acid exchange MLH1 I219V. Conflicting data exist on its effect on the function of the protein and it has recently been reported to cosegregate with refractory UC, suggesting that this alteration may impair mismatch repair activity and thereby contribute to certain forms of UC. We analyzed the MLH1 I219V alteration using in silico and biochemical analyses and assessed its appearance in 67 well-classified UC patients in comparison to 40 healthy individuals. The analyses showed that I219 is a conserved, buried hydrophobic residue, and that I219V is unlikely to abolish MLH1 function but may modulate it. Quantitative biochemical evaluation showed identical stability and activity of the protein. Furthermore, the alteration occurred equally frequently in analyzed patients and healthy volunteers. The MLH1 I219V alteration does not directly contribute to the etiology of UC through an impairment of mismatch repair. A putative linkage disequilibrium of MLH1 I219V with the causative gene(s) of the IBD9 locus is rather distant.

CDld expression on intestinal epithelial cells (IEC) is decreased in inflammatory bowel disease

_Background Based upon current thinking, a potential unifying hypothesis to explain IBD pathogenesis is that IBD represents a dysregulated immune response to some component of the luminal bacteria in a genetically susceptible host. It is therefore important to understand the regulation of molecules which may be involved in orchestrating immune responses to bacteria. CDId is a major histocompatibility complex (MHC) class I-like molecule that is expressed on the apical and lateral membranes of IECs and which is predicted to present hydrophobic antigens, such as bacterial lipids, to T cells. We therefore studied CDld expression in patients with IBD. Methods CDld expression was studied in the intestinal specimens obtained from IBD patients by confocal laser microscopy and biochemical methods. Intestinal tissue specimens were frozen and sectioned in OCT compound, stained with CDId specific antibodies and then visualized using confocal laser microscopy. CDld protein expression was studied biochemically in both grossly affected and normal appearing mucosa of IBD patients using immunoprecipitation with CDld specific antibodies in combination with Western blotting. Results: Confocal microscopy imaging showed that, in contrast to normal human intestine, CDId expression was markedly diminished in IBD tissue. Furthermore, differences were observed in specimens obtained from Ulcerative Colitis (UC) and Crohn's disease patients. Specifically, CDld expression was patchy and fragmented in UC and was minimal to absent in patients with Crohn's disease. These findings were corroborated by biochemical studies which showed that not only was CDId markedly decreased to absent in grossly affected bowel specimens of IBD patients but also that CDld expression was moderately reduced in the normal appearing, grossly unaffected bowel of IBD patients. Conclusion Similar to gp 180, a CD66e homologue involved in activation of CD8 ÷ T cells and which may associate with CDId on IECs, expression of CDld is decreased in IBD. Although the consequences of this event are unknown, it is predicted that IBD intestine contains diminished numbers and/or activity of CDld restricted T cells.