transforming-growth-factor-beta and Colitis--Ulcerative

Enteral nutrition seems to play a significant role in the treatment of both adults and children with active Crohn's disease, and to a lesser degree in the treatment of patients with active ulcerative colitis. The inclusion of some special factors in the enteral nutrition formulas might increase the rate of the efficacy. Actually, enteral nutrition enriched in Transforming Growth Factor-β reduced the activity index and maintained remission in patients with Crohn's disease. In addition, a number of experimental animal studies have shown that colostrum exerts a significantly positive result. Probiotics of a special type and a certain dosage could also reduce the inflammatory process in patients with active ulcerative colitis. Therefore, the addition of these factors in an enteral nutrition formula might increase its effectiveness. Although the use of these formulas is not supported by large clinical trials, it could be argued that their administration in selected cases as an exclusive diet or in combination with the drugs used in patients with inflammatory bowel disease could benefit the patient. In this review, the authors provide an update on the role of enteral nutrition, supplemented with Transforming Growth Factor-β, colostrum, and probiotics in patients with inflammatory bowel disease.

Topics: Adult; Child; Colitis, Ulcerative; Colostrum; Crohn Disease; Dietary Supplements; Enteral Nutrition; Female; Humans; Male; Nutritional Physiological Phenomena; Probiotics; Transforming Growth Factor beta; Treatment Outcome

Crohn's disease (CD) and ulcerative colitis (UC), two chronic and relapsing inflammatory bowel diseases (IBD), are supposed to develop in genetically-predisposed individuals as a result of an excessive immune mucosal response directed against normal components of the gut microbiota. There is also evidence that defects in counter-regulatory mechanisms play a major role in the pathogenesis of IBD. One such a defect involves TGF-β1, a cytokine produced by multiple cells types and able to inhibit pathogenic responses in the gut. In both CD and UC, TGF-β1 is highly produced but unable to signal through the TGF-β receptor-associated Smad pathway and suppress production of inflammatory molecules. Abrogation of TGF-β1 activity has been related to Smad7, an intracellular protein that binds to TGF-β receptor and inhibits TGF-β1-driven Smad-dependent signalling. Indeed, silencing of Smad7 with a specific antisense oligonucleotide restores TGF-β1/Smad signalling, thereby down-regulating inflammatory cytokine production and ameliorating experimental colitis in mice. Altogether these observations led to the development of an oral pharmaceutical compound containing the specific Smad7 antisense oligonucleotide (herein termed GED0301), which seems to be safe and well tolerated in CD patients. In this article we summarize the data supporting the pathogenic role of Smad7 in IBD and discuss the recent results of the use of GED0301 in CD.

Topics: Animals; Clinical Trials, Phase I as Topic; Colitis, Ulcerative; Crohn Disease; Gene Expression Regulation; Humans; Inflammatory Bowel Diseases; Mice; Oligonucleotides; Oligonucleotides, Antisense; Receptors, Transforming Growth Factor beta; Signal Transduction; Smad7 Protein; Transforming Growth Factor beta

It is sometimes difficult but finally possible to distinguish colitis-associated neoplasms from sporadic neoplasms. The frequency of detection of precursor lesions of carcinoma (e.g. dysplasia, intraepithelial neoplasia and adenoma) has increased in recent years, which is most probably due to better endoscopic detection and thus improved histological diagnosis. Carcinogenesis of colitis-associated neoplasms is different from carcinogenesis in sporadic neoplasms because mutations and epigenetic changes are different or may occur at a different point in time. In the present article, these differences will be described and placed in context with carcinogenesis in ulcerative colitis.

Topics: Adenoma; Carcinoma in Situ; Cell Transformation, Neoplastic; Chromosome Aberrations; Colitis, Ulcerative; Colonic Neoplasms; DNA Methylation; DNA Mutational Analysis; Epigenesis, Genetic; Humans; Intestinal Mucosa; Microsatellite Instability; Neoplasm Invasiveness; Neoplasm Staging; Oxidative Stress; Precancerous Conditions; Reactive Oxygen Species; Transforming Growth Factor beta

Topics: Adaptor Proteins, Signal Transducing; Animals; Carrier Proteins; Colitis, Ulcerative; Colorectal Neoplasms; Cyclooxygenase 2; Cytokines; DNA Repair; DNA-Binding Proteins; Genes, APC; Genes, DCC; Genes, p53; Genes, ras; Humans; Inflammation Mediators; Isoenzymes; Membrane Proteins; Mutation; MutL Protein Homolog 1; MutS Homolog 2 Protein; Neoplasm Proteins; Nitric Oxide; Nuclear Proteins; Precancerous Conditions; Prostaglandin-Endoperoxide Synthases; Proto-Oncogene Proteins; Signal Transduction; Transforming Growth Factor beta

Transforming growth factor beta is the protein playing a principal role in the intercellular signalling. The most important functions are: control of cellular growth, differentiation and migration. Moreover it stimulates synthesis of extracellular matrix proteins, modulates immune response and is responsible for angiogenesis, wound formation and tissue reconstruction. All these activities are involved in the development or healing of inflammatory bowel diseases. The role of transforming growth factors beta in the pathogenesis of ulcerative colitis are discussed in this paper.

Topics: Animals; Cell Division; Cell Movement; Colitis, Ulcerative; Gastric Mucosa; Humans; Mesoderm; Neovascularization, Pathologic; Transforming Growth Factor beta

Our study aimed to elucidate the correlation of macrophage (mø) with the inflammatory reaction in ulcerative colitis (UC) and the influence of curcumin (Cur) on mø chemotaxis in mice with UC.. A total of 49 patients with UC (research group; RG) admitted between June 2020 and October 2021 and 56 healthy individuals (control group; CG) who visited concurrently were selected as the study participants. The peripheral blood mononuclear cells (PBMCs) were analyzed, and M1-type/M2-type mø and inflammatory factors (IFs) interleukin (IL)-1, IL-6, IL-10, tumor necrosis factor alpha (TNF-α) and transforming growth factor beta (TGF-β) were detected. In addition, 15 BALB/c mice were purchased and divided into the normal group fed normally, the UC model group established with sodium dextran sulfate (DSS) and the Cur group induced by DSS + Cur feeding. Colon tissue mø was collected from mice to measure mø activity via CCK-8 and to quantify levels of IFs and chemokine CCL2 by polymer chain reaction (PCR)c and Western blotting.. The RG had a higher percentage of peripheral blood M1-type mø and a lower percentage of M2-type mø and M1/M2 mø ratio than the CG (P < .05). In the RG, IL-1, IL-6 and TNF-α all increased and were inversely correlated with the ratio of M1/M2 mø, while IL-10 and TGF-β decreased, with a positive connection with the M1/M2 mø ratio. In the UC model mice, mø activity increased, but the apoptosis rate decreased. mø activity was lower in the Cur group than in the model and normal groups; mø apoptosis in the Cur group was higher than in the model group but lower than in the normal group. In addition, proIFs increased and anti-IFs decreased in the model group, and Cur also ameliorated this process. Finally, CCL2 and MCP-1 levels in the model group were also increased, while those in the Cur group were lower compared with the model group.. In UC, the M1/M2 mø ratio is severely misadjusted, activation of M1-type mø is enhanced and pro-IFs are released in large quantities. Cur can ameliorate the abnormal activation of mø in mice with UC, inhibit mø chemotaxis and alleviate the inflammatory reaction, which may make it a new option for UC treatment in the future.

Topics: Animals; Chemotaxis; Colitis, Ulcerative; Curcumin; Disease Models, Animal; Inflammation; Interleukin-10; Interleukin-6; Leukocytes, Mononuclear; Macrophages; Mice; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Ulcerative colitis (UC) is a chronic mucosal inflammation of the large intestine that mostly affects the rectum and colon. The absence of safe and effective therapeutic agents encourages the discovery of novel therapeutic agents to effectively treat UC and its complications. The purpose of this research was to examine the protective impact of Eicosapentaenoic acid (EPA) in rats with UC induced by acetic acid (AA).. AA (2 ml, 3 % v/v) was injected intrarectally to cause UC. Before administering AA, EPA (300 and 1000 mg/kg) was given orally for 28 days.. EPA inhibited AA-induced UC by enhancing colonic histopathological changes like inflammation, goblet cell loss, glandular hyperplasia and mucosal ulceration, concomitant with a reduction in colon weight, colon weight/length ratio, C-reactive protein (CRP), and serum lactate dehydrogenase (LDH). EPA also effectively restored the imbalance between oxidants and antioxidants caused by AA. In addition, EPA increased the levels of trefoil factor-3 (TFF-3) and glucagon-like peptide-1 (GLP-1), while significantly reducing the expression of nuclear factor kappa B (NF-κB), interferon-γ (IFN-γ), and interleukin-6 (IL-6), transforming growth factor-1(TGF-β1), and phosphorylated epidermal growth factor receptor (P-EGFR), phosphatidylinositol-3-kinase (PI3K) and protein kinase B (AKT) expression in colonic tissues.. EPA inhibited AA-induced UC in rats by modulating the TGF-β/P-EGFR and NF-κB inflammatory pathways, regulating the oxidant/antioxidant balance, and enhancing the colon barrier integrity.

Topics: Acetic Acid; Animals; Antioxidants; Colitis, Ulcerative; Colon; Eicosapentaenoic Acid; ErbB Receptors; Inflammation; NF-kappa B; Rats; Signal Transduction; Transforming Growth Factor beta

Ulcerative colitis (UC), limited to the colon's innermost lining, has become a global health problem. Immunomodulatory and monoclonal antibodies are used to treat UC despite their side effects and limitations. Phenytoin is used to heal wounds owing to its effects on growth factors, collagen, and extracellular matrix synthesis. This study aimed to evaluate the effect of topical phenytoin administration in UC. Phenytoin was administered in two doses during the treatment. Eighty male Wistar rats (230-280 g) were divided randomly into ten groups of sham, control, hydrocortisone, phenytoin 1%, and 3% groups in 6- or 12-day treatment protocols. The UC model was induced by the administration of acetic acid 4% into the colon. Animals were killed on the 7th and 13th postoperative days. The main outcome measures included body weight loss, microscopic score, and ulcer index measured using specific criteria. Growth factors were measured by western blotting. Results illustrated that body weight loss was reversed in the treatment groups. Ulcer index had decreased on 6- and 12-day treatment protocols. Microscopic scores in 6-day enema treatment significantly decreased compared to the control groups. Transforming growth factor-beta (TGFβ) significantly increased in a time-dependent manner and platelet-derived growth factor (PDGF) and vascular endothelial growth factor (VEGF) significantly increased in a time- and dose-dependent manner in phenytoin 1% and 3% in the 6- and 12-day protocols. Phenytoin dose- and time-dependently reversed weight loss. In addition, histopathological parameters included microscopic scores, and the ulcer index was decreased through the induction of growth factors TGFβ, PDGF, and VEGF and consequently accelerated ulcer healing.

Topics: Acetic Acid; Animals; Colitis, Ulcerative; Male; Phenytoin; Platelet-Derived Growth Factor; Rats; Rats, Wistar; Transforming Growth Factor beta; Transforming Growth Factors; Vascular Endothelial Growth Factor A

Aberrant TGF‑β/Smad7 signaling has been reported to be an important mechanism underlying the pathogenesis of ulcerative colitis. Therefore, the present study aimed to investigate the effects of a number of potential anti‑colitis agents on intestinal epithelial permeability and the TGF‑β/Smad7 signaling pathway in an experimental model of colitis. A mouse model of colitis was first established before anti‑TNF‑α and 5‑aminosalicyclic acid (5‑ASA) were administered intraperitoneally and orally, respectively. Myeloperoxidase (MPO) activity, histological index (HI) of the colon and the disease activity index (DAI) scores were then detected in each mouse. Transmission electron microscopy (TEM), immunohistochemical and functional tests, including Evans blue (EB) and FITC‑dextran (FD‑4) staining, were used to evaluate intestinal mucosal permeability. The expression of epithelial phenotype markers E‑cadherin, occludin, zona occludens (ZO‑1), TGF‑β and Smad7 were measured. In addition, epithelial myosin light chain kinase (MLCK) expression and activity were measured. Anti‑TNF‑α and 5‑ASA treatments was both found to effectively reduce the DAI score and HI, whilst decreasing colonic MPO activity, plasma levels of FD‑4 and EB permeation of the intestine. Furthermore, anti‑TNF‑α and 5‑ASA treatments decreased MLCK expression and activity, reduced the expression of Smad7 in the small intestine epithelium, but increased the expression of TGF‑β. In mice with colitis, TEM revealed partial epithelial injury in the ileum, where the number of intercellular tight junctions and the expression levels of E‑cadherin, ZO‑1 and occludin were decreased, all of which were alleviated by anti‑TNF‑α and 5‑ASA treatment. In conclusion, anti‑TNF‑α and 5‑ASA both exerted protective effects on intestinal epithelial permeability in an experimental mouse model of colitis. The underlying mechanism may be mediated at least in part by the increase in TGF‑β expression and/or the reduction in Smad7 expression, which can inhibit epithelial MLCK activity and in turn reduce mucosal permeability during the pathogenesis of ulcerative colitis.

Topics: Animals; Cadherins; Colitis, Ulcerative; Colon; Dextran Sulfate; Disease Models, Animal; Female; Intestinal Mucosa; Male; Mesalamine; Mice, Inbred C57BL; Myosin-Light-Chain Kinase; Occludin; Peroxidase; Severity of Illness Index; Signal Transduction; Smad7 Protein; Tight Junctions; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Zonula Occludens-1 Protein

In clinical practice, colorectal cancer (CRC) is difficult to distinguish from ulcerative colitis and colon polyps. Practical markers are useful for diagnosing and treating patients with CRC. Carcinoembryonic antigen (CEA) is a biomarker for diagnosing patients with CRC. However, the diagnostic sensitivity and specificity of CEA are not high. Interleukin (IL)-10, IL-17A, tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), and transforming growth factor beta (TGF-β) are assumed to be closely related to the occurrence and development of human cancer. Some have been used as diagnostic markers in CRC. It remains unclear whether cytokines in combination with CEA could be used as biomarkers for the diagnosis of CRC. Serum levels of IL-10, IL-17, TNF-α, IFN-γ, and TGF-β in patients with CRC, ulcerative colitis, colonic polyps, stomach cancer, and healthy controls were measured by enzyme-linked immunosorbent assay. The serum level of CEA was detected using electrochemiluminescence. The value of the cytokines combined with CEA as a biomarker panel for the diagnosis of CRC was assessed. CEA, IL-10, IL-17A, TNF-α, and TGF-β levels were significantly increased in CRC. CEA displayed a higher specificity than the other cytokines. IL-17A, TNF-α, and TGF-β displayed higher sensitivities than CEA, IL-10, and IFN-γ in the diagnosis of CRC. The combination of serum CEA, IL-17A, and TNF-α achieved higher diagnostic efficacy for CRC (area under the curve = 0.935). The combination of CEA, IL-17, and TNF-α has better diagnostic efficacy than CEA alone in CRC. A panel containing IL-17A, TNF-α, and CEA could be a promising molecular biomarker panel to diagnostically differentiate CRC from ulcerative colitis, colon polyps, and stomach cancer.

Topics: Biomarkers, Tumor; Carcinoembryonic Antigen; Colitis, Ulcerative; Colonic Polyps; Colorectal Neoplasms; Cytokines; Humans; Interferon-gamma; Interleukin-10; Interleukin-17; Stomach Neoplasms; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Although big data from transcriptomic analyses have helped transform our understanding of inflammatory bowel disease (IBD), they remain underexploited. We hypothesized that the application of machine learning using lasso regression to transcriptomic data from IBD patients and controls can help identify previously overlooked genes. Transcriptomic data provided by Ostrowski et al. (ENA PRJEB28822) were subjected to a two-stage process of feature selection to discriminate between IBD and controls. First, a principal component analysis was used for dimensionality reduction. Second, the least absolute shrinkage and selection operator (lasso) regression was employed to identify genes potentially involved in the pathobiology of IBD. The study included data from 294 participants: 100 with ulcerative colitis (48 adults and 52 children), 99 with Crohn's disease (45 adults and 54 children), and 95 controls (46 adults and 49 children). IBD patients presented a wide range of disease severity. Lasso regression preceded by principal component analysis successfully selected interesting features in the IBD transcriptomic data and yielded 12 models. The models achieved high discriminatory value (range of the area under the receiver operating characteristic curve 0.61-0.95) and identified over 100 genes as potentially associated with IBD.

Topics: Adult; Child; Colitis, Ulcerative; Humans; Inflammatory Bowel Diseases; Machine Learning; Transcriptome; Transforming Growth Factor beta

Ulcerative colitis (UC) is an inflammatory disease, and studies have suggested a role for TGF-β signalling pathway in the pathogenesis of UC. In the present study, we evaluated expression of TGF-β signalling genes and their regulatory microRNAs in patients with UC and control subjects. The expression of TGF-β1, SMAD2, SMAD3, miR-21, miR-101, miR-433, and miR-590 were evaluated using real-time PCR in biopsy samples of the patients and controls. Results showed increased expression of TGF-β1 and SMAD3 in the patients compared to controls. In addition, miR-21 and miR-433 were found to be higher in the patients compared to controls; however, miR-590 was found to be lower. Moreover, miR-433 was demonstrated to have positive correlation with SMAD3 and TGF-β while miR-21 was positively correlated with TGF-β1. MiR-590 was negatively correlated with SMAD2 and SMAD3. Results of the present study suggested a role for TGF-β signalling pathway related microRNAs in pathogenesis of UC.

Topics: Colitis, Ulcerative; Humans; MicroRNAs; Signal Transduction; Smad3 Protein; Transforming Growth Factor beta; Transforming Growth Factor beta1

Topics: Akkermansia; Animals; Colitis, Ulcerative; Colon; Dextran Sulfate; Fecal Microbiota Transplantation; Gastrointestinal Microbiome; Helicobacter; Interleukins; Male; Mice; Mice, Inbred C57BL; T-Lymphocytes; Transforming Growth Factor beta

Myricetin is a bioactive compound in many edible plants with anti-inflammatory and anticarcinogenic activity. The current study aimed to determine the protective effects and mechanism of myricetin against ulcerative colitis (UC).. Myricetin was orally administered at doses of 40 and 80 mg/kg to C57BL/6 mice with UC induced using dextran sulfate sodium. The disease-associated index and colon length were determined at the end of the experiment, the proportion of Treg, Th1 and Th17 was analysed by cytometry, and cytokines were detected using ELISA.. Myricetin (80 mg/kg) ameliorated the severity of inflammation in acute UC and significantly improved the condition. Myricetin (80 mg/kg) elevated the levels of IL-10 and transforming growth factor β. In addition, the proportion of regulatory T cells significantly increased in mice in the myricetin treatment group.. Taking together, these results suggest that myricetin exhibits significant protective effects against UC and it could be used as a potential treatment for UC.

Topics: Animals; Anti-Inflammatory Agents; Colitis, Ulcerative; Colon; Dextran Sulfate; Disease Models, Animal; Female; Flavonoids; Interleukin-10; Mice, Inbred C57BL; T-Lymphocytes, Regulatory; Th1 Cells; Th17 Cells; Transforming Growth Factor beta

Ulcerative colitis (UC) and Crohn's disease (CD) are two major forms of inflammatory bowel disease (IBD), which is an inflammatory disease. Studies have shown that adipose tissue and inflammation play important roles in the pathogenesis of IBD. C1q/TNF-related protein-3 (CTRP3) is a newly discovered adipokine playing a substantial role during inflammatory process, and for the first time in the present study, serum levels of this adipokine were measured in the UC and CD patients. This case-control study included 70 control, 50 UC, and 50 CD patients who were diagnosed by standard criteria. Serum levels of adiponectin, IL-6, TNF-α, TGF-β, and CTRP3 were evaluated using ELISA kits. Serum levels of IL-6, TNF-α, and TGF-β elevated in the UC and CD patients compared with the controls while adiponectin and CTRP3 diminished in the patient's groups compared with the control. Furthermore, decrease in CTRP3 serum levels was associated with the risk of UC and CD diseases. Moreover, CTRP3 indicated negative correlation with BMI, FBS, insulin, homeostasis model assessment of insulin resistance, IL-6, TNF-α, and TGF-β and also a positive correlation with adiponectin in both the UC and CD patients. For the first time, the present study demonstrated lower levels of CTRP3 in the UC and CD patients. Decreased serum levels of CTRP3 and its inverse relationship with inflammatory cytokines and TGF-β levels suggested a possible role for CTRP3 in the pathogenesis of UC and CD diseases.

Topics: Adipokines; Adiponectin; Adult; Colitis, Ulcerative; Crohn Disease; Cytokines; Female; Humans; Inflammatory Bowel Diseases; Insulin; Insulin Resistance; Interleukin-6; Male; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Tumor Necrosis Factors

Diagnostic markers for distinguishing between Crohn disease (CD) and ulcerative colitis (UC) remain elusive. We studied whether methylation marks across the promoters of the transforming growth factor beta 1 (TGFβ1) and interleukin-6 genes have diagnostic utility.. A case-control study was carried out. Cases were treatment-naïve, diagnosed before age 20, and recruited from 3 pediatric gastroenterology clinics across Canada. Control patients did not have inflammatory bowel disease and were recruited from orthopedic clinics within the same hospitals as the gastroenterology clinics. Patient DNA from peripheral blood was processed to identify methylation sites (CpG) across the promoter regions of the TGFβ1 and interleukin-6 genes. After initial nonparametric univariate analyses, multivariate logistic regression models were fit. Models with the best fit (Akaike information criteria) and strongest discriminatory capabilities (area under the curve [AUC]) were identified, and P values were adjusted for multiple comparisons using the false discovery rate method.. A total of 67 CD, 31 UC, and 43 control patients were included. The age distribution of the 3 groups was similar. Most CD patients had ileocolonic disease (44.8%) and inflammatory disease (88.1%). Most UC patients had extensive (71%) and moderate disease (51.6%). Logistic regression analysis revealed the following: 14 TGFβ1 CpG sites discriminated between CD and control patients (AUC = 0.94), 9 TGFβ1 CpG sites discriminated between UC and control patients (AUC = 0.99), 3 TGFβ1 CpG sites discriminated between CD and UC (AUC = 0.81), and 6 TGFβ1 CpG sites distinguished colonic CD from UC (AUC = 0.91).. We found that CpG methylation in the promoter of the TGFβ1 gene has high discriminative power for identifying CD and UC and could serve as an important diagnostic marker.

Topics: Adolescent; Area Under Curve; Biomarkers; Canada; Case-Control Studies; Child; Colitis, Ulcerative; CpG Islands; Crohn Disease; Diagnosis, Differential; Female; Humans; Interleukin-6; Logistic Models; Male; Methylation; Transforming Growth Factor beta; Young Adult

To investigate the effect of decitabine on the regulation of intestinal barrier function in mice with inflammatory bowel disease, an experimental model of colitis was established via drinking water with dextran sulfate sodium (DSS). Hematoxylin and eosin staining was used to observe the pathological changes of the colon. Cytokine production was measured by an ELISA assay. Flow cytometry was used to measure the level of regulatory T cells. Immunofluorescence, immunohistochemistry and western blot analyses detected the protein expression and distribution in colon tissue. Following the administration of decitabine, the symptoms of intestinal inflammation in the mice were significantly relieved; the expression of IL‑17 was decreased, and the levels of TGF‑β and IL‑10 were increased. In addition, the induction of forkhead box P3 (Foxp3) in naive T cells increased the proportion of CD4+ Foxp3+ T cells in CD4+ T cells. Furthermore, decitabine increased the levels of zonular occludens‑1 and occludin, and inhibited the phosphorylation of ERK1/2, JNK and p38. In conclusion, the present study suggested that decitabine could alleviate DSS‑induced impaired colon barrier and the weight loss, mucus and bloody stools in mice by releasing the inhibitory factor IL‑10, reducing the pro‑inflammatory factor IL‑17, activating CD4+ Foxp3+ T cells and inhibiting the activation of the MAPK pathway.

Topics: Animals; Blotting, Western; CD4-Positive T-Lymphocytes; Colitis, Ulcerative; Colon; Decitabine; Dextran Sulfate; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Forkhead Transcription Factors; Immunohistochemistry; Interleukin-10; Interleukin-17; Male; Mice; Mice, Inbred BALB C; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

Ulcerative colitis is one of the IBD which cause a chronic intestinal inflammation and dysfunctional of the mucosal barrier. For now, the incident of UC was steadily increased all over the world. It has become a novel independent risk factor of several severe diseases especially colon-rectal cancer. However, the etiology of UC was still obscure. Previous studies show that high-fat diet contributed to the pathogenesis of immune system dysregulation, and farnesoid X receptor (FXR) was also implicated in the pathogenesis of various inflammatory symptoms. Yet, their inner roles in the pathogenesis of UC have not been mentioned. In this study, we aim to investigate the role of FXR in UC. High-fat diet (HFD) promotes the progression of DSS-induced UC, shows an increasing secretion of bile acid in serum, and leads to a downregulation of FXR target genes (FXR

Topics: Animals; Caco-2 Cells; Colitis, Ulcerative; Dextran Sulfate; Diet, High-Fat; Disease Models, Animal; Disease Progression; Down-Regulation; Humans; Male; Mice, Inbred C57BL; Receptors, Cytoplasmic and Nuclear; Signal Transduction; Transforming Growth Factor beta; Up-Regulation

Topics: Adolescent; Child; Colitis, Ulcerative; Crohn Disease; Enteral Nutrition; Female; Growth Disorders; Hematologic Tests; Humans; Male; Malnutrition; Remission Induction; Retrospective Studies; Transforming Growth Factor beta

A healed intestinal mucosa is the aim of therapy in acute ulcerative colitis (UC). Disruption of mucosal wound healing may lead to severe complications including intestinal fibrosis. This study examined mucosal gene expression in the healing process of acute UC with a special focus on known mediators of fibrosis.. Endoscopic biopsies from patients with acute, moderate to severe UC were analyzed with a quantitative polymerase chain reaction array for 84 genes involved in fibrosis pathways. All patients were treated with infliximab (anti- tumor necrosis factor). Biopsies were taken before therapy and when disease remission was reached, defined as a Mayo score of ≤2, with an endoscopic subscore of 0 or 1. A healthy control group was included. Immunostaining of matrix metallopeptidase 9 and smooth muscle actin was performed.. Mucosal biopsies from acute UC (n = 28), remission UC (n = 28), and healthy controls (n = 13) were analyzed. Fibrosis and extracellular matrix-associated genes were upregulated in the endoscopically healed UC mucosa vs controls, with collagen type III alpha 1 chain, actin alpha 2, lysyl oxidase, TIMP metallopeptidase inhibitor 3, and caveolin 1 uniquely showing no overlap with acute disease. Pro- and antifibrotic mediators (interleukin [IL]13 receptor subunit alpha 2, IL1B, IL10, tumor necrosis factor, snail family transcriptional repressor 1, and C-C motif chemokine ligand 2) were upregulated in both acute and healed UC compared with controls. An attenuated pattern of the canonical transforming growth factor beta (TGFB) pathway was observed in acute UC and to a lesser extent in the healed mucosa, except for TGFB2, which was enhanced.. The endoscopically healed mucosa of UC showed a persisting dysregulation of fibrosis-associated mediators compared with controls, including extracellular matrix remodeling, profibrotic cytokines, and TGFB signaling pathways.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Biopsy; Case-Control Studies; Colitis, Ulcerative; Colon; Colonoscopy; Cytokines; Extracellular Matrix; Female; Fibrosis; Gastrointestinal Agents; Gene Expression Profiling; Humans; Infliximab; Intestinal Mucosa; Male; Middle Aged; Severity of Illness Index; Signal Transduction; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Wound Healing; Young Adult

Ulcerative colitis (UC) is one of the inflammatory diseases of the gut with frequent bloody diarrhea leads to increased rates of anemia. Evidences indicate the immunomodulation disorders in the response to intestinal microbiota in UC. Although sugarcane molasses, rich in necessary minerals and vitamins, could be a good support nutrient but its effect on immune system of UC patients is unknown. To determine how the immune system of UC patients responds to molasses this study was planned. Bifidobacterium lactis were cultivated on MRS broth. PBMCs of 12 UC patients were separated by Ficoll-Hypaque centrifugation and co-cultured with different concentrations of UV killed bacteria and/or molasses in RPMI-1640 plus 10 % FCS. The gene expression of FoxP3 was measured by real-time PCR. TGF-β and TNF-α were measured in supernatant of PBMCs by ELISA. Sugarcane molasses and B. lactis significantly augmented TGF-β compared to control (p < 0.01 and p < 0.001 respectively). The secretion levels of TGF-β by B. lactis plus molasses compared to B. lactis stimulated PBMCs was significantly higher (p < 0.05) but the level of TNF-α by PBMCs after 2/4/12 h incubation with B. lactis plus molasses compared to B. lactis alone was not changed (p > 0.2). The level of FOXP3 expression after treatment with molasses was increased significantly (p < 0.05). These data show that if sugarcane molasses added to B. lactis, not only do not increase the pro-inflammatory cytokine, TNF-α, but also augments the anti-inflammatory cytokine, TGF-β by PBMCs. Therefore, these results pave the way for further investigation to show sugarcane molasses as a safe support to compensate the lost nutrients in UC patients.

Topics: Adult; Bifidobacterium animalis; Colitis, Ulcerative; Female; Forkhead Transcription Factors; Humans; Leukocytes, Mononuclear; Male; Molasses; Saccharum; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

MicroRNAs [miRNAs] have emerged as important regulators in inflammatory bowel disease [IBD]. This study investigated differential expression of miRNAs across clinical phenotypes in a well-characterized cohort of IBD patients and healthy controls [HCs].. A cohort of Crohn's disease [CD] and ulcerative colitis [UC] patients and HCs was prospectively accrued. Total RNA was extracted from peripheral blood mononuclear cells for all subjects. miRNA expression was measured using NanoString technologies. The subjects were stratified according to disease activity and location. Statistical significance was assessed per miRNA across outcomes and corrected for multiple testing. miRNA regulation of transcription of important results was confirmed in vitro by a dual luciferase reporter assay and autophagy function was evaluated using immunofluorescence imaging of LC3 puncta in HeLa cells.. In total, 120 subjects were enrolled. Seventy-four miRNAs were differentially expressed across CD, UC and HCs. Comparing quiescent CD [CDq] with HCs we found ten miRNAs upregulated in CDq. When comparing colonic CD [CCD] to UC, seven miRNAs were upregulated in CCD. The most differentially expressed miRNA in CCD vs UC was miR-874-3p, and we showed its possible utility as a biomarker of differential diagnosis. We showed miR-874-3p targets ATG16L1 and reduces its expression in vitro. An miR-874-3p mimic dysregulates autophagy by a reduction of LC3 in vitro.. We identified unique miRNA signatures expressed in distinct IBD phenotypes. These associations highlight pathways dysregulated by aberrant miRNA expression, revealing possible mechanisms underlying the pathophysiology of IBD, but also suggest a cluster of miRNAs as readily accessible biomarkers to aid in differential diagnosis.

Topics: Adult; Aged; Autophagy; Autophagy-Related Proteins; Biomarkers; Case-Control Studies; Colitis, Ulcerative; Colon; Crohn Disease; Diagnosis, Differential; Female; HeLa Cells; Humans; Leukocytes, Mononuclear; Male; MicroRNAs; Middle Aged; Prospective Studies; Severity of Illness Index; Signal Transduction; Smad3 Protein; Transforming Growth Factor beta; Up-Regulation; Young Adult

T lymphocytes represent an important part of adaptive immune system undertaking different functions to regulate immune responses. CD4+ T cells are the most important activator cells in inflammatory conditions. Depending on the type of induced cells and inflamed sites, expression and activity of different subtypes of helper T cells are changed. Recent studies have confirmed the existence of a new subset of helper T lymphocytes called Th9. Naive T cells can differentiate into Th9 subtypes if they are exposed simultaneously by interleukin (IL) 4 and transforming growth factor β and also secondary activation of a complicated network of transcription factors such as interferon regulatory factor 4 (IRF4) and Smads which are essential for adequate induction of this phenotype. Th9 cells specifically produce interleukin 9 and their probable roles in promoting intestinal inflammation are being investigated in human subjects and experimental models of ulcerative colitis (UC). Recently, infiltration of Th9 cells, overexpression of IL-9, and certain genes associated with Th9 differentiation have been demonstrated in inflammatory microenvironment of UC. Intestinal oversecretion of IL-9 protein is likely to break down epithelial barriers and compromise tolerance to certain commensal microorganisms which leads to inflammation. Th9 pathogenicity has not yet been adequately explored in UC and they are far from being considered as inflammatory cells in this milieu, therefore precise understanding the role of these newly identified cells in particular their potential role in gut pathogenesis may enable us to develop novel therapeutic approaches for inflammatory bowel disease. So, this article tries to discuss the latest knowledge on the above-mentioned field.

Topics: Cell Differentiation; Colitis, Ulcerative; Colon; Gastrointestinal Microbiome; Humans; Immune Tolerance; Interferon Regulatory Factors; Interleukin-4; Interleukin-9; Intestinal Mucosa; Signal Transduction; Smad Proteins; T-Lymphocyte Subsets; T-Lymphocytes, Helper-Inducer; Transforming Growth Factor beta

Cholera toxin B subunit (CTB) is a component of a licensed oral cholera vaccine. However, CTB has pleiotropic immunomodulatory effects whose impacts on the gut are not fully understood. Here, we found that oral administration in mice of a plant-made recombinant CTB (CTBp) significantly increased several immune cell populations in the colon lamina propria. Global gene expression analysis revealed that CTBp had more pronounced impacts on the colon than the small intestine, with significant activation of TGFβ-mediated pathways in the colon epithelium. The clinical relevance of CTBp-induced impacts on colonic mucosa was examined. In a human colon epithelial model using Caco2 cells, CTBp, but not the non-GM1-binding mutant G33D-CTBp, induced TGFβ-mediated wound healing. In a dextran sodium sulfate (DSS) acute colitis mouse model, oral administration of CTBp protected against colon mucosal damage as manifested by mitigated body weight loss, decreased histopathological scores, and blunted escalation of inflammatory cytokine levels while inducing wound healing-related genes. Furthermore, biweekly oral administration of CTBp significantly reduced disease severity and tumorigenesis in the azoxymethane/DSS model of ulcerative colitis and colon cancer. Altogether, these results demonstrate CTBp's ability to enhance mucosal healing in the colon, highlighting its potential application in ulcerative colitis therapy besides cholera vaccination.

Topics: Administration, Oral; Animals; Azoxymethane; Caco-2 Cells; Cholera; Cholera Toxin; Cholera Vaccines; Colitis, Ulcerative; Colon; Colonic Neoplasms; Dextran Sulfate; Disease Models, Animal; Female; Humans; Mice; Mice, Inbred C57BL; Mucous Membrane; Signal Transduction; Transforming Growth Factor beta; Wound Healing

A CD4+CD25- regulatory T cell population expressing the surface TGF-β in its latent form LAP+ [latency associated peptide] cells was proved to be protective in experimental colitis and to be suppressive of human peripheral blood [PB] T proliferation. We investigated the frequency and function of lamina propria [LP] CD4+LAP+ T cells in inflammatory bowel disease [IBD] patients.. Specimens from patients undergoing colonoscopy or bowel resection for IBD and colonic cancer were used as source of lamina propria mononuclear cells [LPMC]. The ulcerative colitis [UC] group was divided according to endoscopic activity evaluated with modified Baron Score. IL-17, IFN-γ, IL-10, LAP, and Foxp3 expression in CD3+CD8- [CD4] or CD3+/CD4+ gated cell population was assessed by immunofluorescence. The ability of FACS-sorted LP CD3+CD8-[CD4] LAP+CD25- to inhibit stimulated autologous PB CD3+CD8-[CD4] LAP- CD25- cells proliferation was assessed.. LP CD4LAP+ cells were significantly increased, when compared with controls, in active UC patients and not in Crohn's disease patients. The majority of LP CD4+LAP+ cells were Foxp3-. The percentage of IL-17+ cells in LP CD3+CD8-[CD4] LAP+ cells was significantly higher in active UC patients when compared with controls. LP CD3+CD8-[CD4]LAP+CD25- isolated from UC patients showed reduced or no ability to inhibit autologous PB CD3+CD8-[CD4]LAP-CD25- cell proliferation when compared with controls. Removal of IL-17+ cells from LP CD3+CD8-[CD4] LAP+ cells increases their suppressive ability.. The percentage of LP CD4LAP+ cells is increased in active UC, showing reduced suppressor activity due to their increased proportion of intracellular IL-17 expression.

Topics: Adult; Aged; Biomarkers; Case-Control Studies; CD4 Lymphocyte Count; Cell Proliferation; Colitis, Ulcerative; Colon; Crohn Disease; Female; Humans; Ileum; Interleukin-17; Intestinal Mucosa; Male; Middle Aged; Peptides; Protein Precursors; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

The inflammatory state associated with Crohn's disease (CD) and ulcerative colitis (UC) remains incompletely defined. To understand more clearly the extracellular milieu associated with inflammatory bowel disease (IBD), we employed a bioassay whereby plasma of treatment naive paediatric IBD patients (n = 22 CD, n = 15 UC) and unrelated healthy controls (uHC, n = 10) were used to induce transcriptional responses in a healthy leucocyte population. After culture, gene expression was measured comprehensively with microarrays and analysed. Relative to uHC, plasma of CD and UC patients induced distinct responses consisting, respectively, of 985 and 895 regulated transcripts [|log2 ratio| ≥ 0·5 (1·4-fold); false discovery rates (FDR) ≤ 0·01]. The CD:uHC and UC:uHC signatures shared a non-random, commonly regulated, intersection of 656 transcripts (χ(2)  = P < 0·001) and were highly correlative [Pearson's correlation coefficient = 0·96, 95% confidence interval (CI) = 0.96, 0.97]. Despite sharing common genetic susceptibility loci, the IBD signature correlated negatively with that driven by plasma of type 1 diabetes (T1D) patients (Pearson's correlation coefficient = -0·51). Ontological analyses revealed the presence of an immunoregulatory plasma milieu in IBD, as transcripts for cytokines/chemokines, receptors and signalling molecules consistent with immune activation were under-expressed relative to uHC and T1D plasma. Multiplex enzyme-linked immunosorbent assay (ELISA) and receptor blockade studies confirmed transforming growth factor (TGF)-β and interleukin (IL)-10 as contributors to the IBD signature. Analysis of CD patient signatures detected a subset of transcripts associated with responsiveness to 6-mercaptopurine treatment. Through plasma-induced signature analysis, we have defined a unique, partially TGF-β/IL-10-dependent immunoregulatory signature associated with IBD that may prove useful in predicting therapeutic responsiveness.

Topics: Adolescent; Blood Proteins; Child; Child, Preschool; Colitis, Ulcerative; Crohn Disease; Diabetes Mellitus, Type 1; Female; Healthy Volunteers; Humans; Immunologic Factors; Interleukin-10; Leukocytes, Mononuclear; Male; Primary Cell Culture; Protein Array Analysis; RNA, Messenger; Transcriptome; Transforming Growth Factor beta

To investigate IL-1α, IL-1β, IL-1R , IL-4RA, TGF-β, TNF-α and IFN-γ, genes polymorphism in Turkish patients with ucerative colitis (UC).. An analysis was carried out at Trabzon Karadeniz Technical University Medicine Faculty Gastroenterology polyclinics between March 2005 and May 2011 on 51 patient with UC (cases) and 100 healthy individuals (controls). PCR-SSP and cytokine gene panel (Helderberg kits) based techniques for analysis of gene polymorphisms were used.. Changes in allelic frequencies of each of the investigated eight cytokine genes polymorphisms in patient with ulcerative colitis were found. Among the allelic genes analyzed here, the highest statistically significant change was observed in the position TNF-α -308 G/A (339.7%). The following increases were observed in IL-IR mspa T/C variation (179.4%), IFN-γ 5644A/T variation (77.4%), and in IL-1β -511T/C SNPs (35.9% ). In other analyzed genes, allelic changes were found to be decreasing for TGF- β codon10C / T (-71.9%), IL4RA + 1902G / A (-47.3 %), and for IL- 1α -889T / C (-37.7%). The lowest negative change (-25.9%) was observed in the allele frequency in IL- 1β 3962T / C (p les than 0.000). In addition, there were changes in genotypic frequencies investigated seven gene polymophic site and only one of cytokine gene IL-1β 3962TT/TC/CC was not changed.. Genes polymorphism is not itself the only determining factor for clinical diagnoses. However, it can be used in the clinical diagnosis of UC in order to determine the low level or high level variations in cytokine gene polymorphisms.

Topics: Adolescent; Adult; Aged; Colitis, Ulcerative; Cytokines; Female; Gene Frequency; Humans; Interferon-gamma; Interleukins; Male; Middle Aged; Polymorphism, Single Nucleotide; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Turkey; Young Adult

Much of our understanding of gut-microbial interactions has come from mouse models. Intestinal immunity is complex and a combination of host genetics and environmental factors play a significant role in regulating intestinal immunity. Due to this complexity, no mouse model to date gives a complete and accurate representation of human intestinal diseases, such as inflammatory bowel diseases. However, intestinal tissue from patients undergoing bowel resection reflects a condition of severe disease that has failed treatment; hence a more dynamic perspective of varying inflammatory states in IBD could be obtained through the analyses of pinch biopsy material. Here we describe our protocol for analyzing mucosal pinch biopsies collected predominantly during colonoscopies. We have optimized flow cytometry panels to analyze up to 8 cytokines produced by CD4+ and CD8+ cells, as well as for characterizing nuclear proteins and transcription factors such as Ki67 and Foxp3. Furthermore, we have optimized approaches to analyze the production of cytokines, including TGF-beta from direct ex vivo cultures of pinch biopsies and LPMCs isolated from biopsies. These approaches are part of our workflow to try and understand the role of the gut microbiota in complex and dynamic human intestinal diseases.

Topics: Biopsy; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Colitis, Ulcerative; Colon; Colonoscopy; Crohn Disease; Cytokines; Forkhead Transcription Factors; Humans; Intestinal Mucosa; Ki-67 Antigen; Microbiota; Transforming Growth Factor beta

Regulatory T (Treg) cells expressing the transcription factor forkhead box P3 (FoxP3) control immune responses and prevent autoimmunity. Treatment with glucocorticoids (GCs) has been shown to increase Treg cell frequency, but the mechanisms of their action on Treg cell induction are largely unknown. Here, we report that glucocorticoid-induced leucine zipper (GILZ), a protein induced by GCs, promotes Treg cell production. In mice, GILZ overexpression causes an increase in Treg cell number, whereas GILZ deficiency results in impaired generation of peripheral Treg cells (pTreg), associated with increased spontaneous and experimental intestinal inflammation. Mechanistically, we found that GILZ is required for GCs to cooperate with TGF-β in FoxP3 induction, while it enhances TGF-β signaling by binding to and promoting Smad2 phosphorylation and activation of FoxP3 expression. Thus, our results establish an essential GILZ-mediated link between the anti-inflammatory action of GCs and the regulation of TGF-β-dependent pTreg production.

Topics: Animals; Colitis, Ulcerative; Forkhead Transcription Factors; Glucocorticoids; Mice; Mice, Inbred C57BL; Signal Transduction; T-Lymphocytes, Regulatory; Transcription Factors; Transforming Growth Factor beta

Patients with ulcerative colitis have increased risk of colorectal carcinoma, but little is known about how peritoneal macrophages are involved in ulcerative colitis-associated carcinogenesis. We investigated the alteration of peritoneal macrophages and M1/M2 subpopulations during ulcerative colitis-associated carcinogenesis.. Expression and functional changes in peritoneal macrophages and M1/M2 subpopulations were investigated by histopathology, flow cytometry, immunofluorescence, cytokines expression by ELISA and QRT-PCR in an azoxymethane (AOM)- and dextran sodium sulfate (DSS)-induced chemical colitis-associated carcinoma mouse model using male Crj:CD-1 (ICR) mice.. Striking evidence observed in histopathology, flow cytometry, cytokine detection, and gene expression analysis all revealed that inflammation-associated cytokines (IL-1β, IL-10, IL-12, IL-6, TNF-α) and migration/invasion-associated factors (G-CSF, GM-CSF, CXCR4, VEGF, TGF-β, ICAM-1) induced by peritoneal M2 macrophages increased significantly during the progression from inflammatory hyperplasia to carcinoma and metastasis. Similar functional changes occurred during peritoneal metastasis in M1 macrophages without changed polarization.. These results suggested that peritoneal M2 macrophages played a critical role in ulcerative colitis-associated carcinogenesis, including unbalanced pro-inflammatory and anti-inflammatory axis and enhanced expression of migration/invasion-associated factors. Furthermore, functional changes of M1 macrophages occurred without changed polarization during carcinogenesis and metastasis.

Topics: Animals; Colitis, Ulcerative; Colorectal Neoplasms; Cytokines; Disease Models, Animal; Granulocyte Colony-Stimulating Factor; Granulocyte-Macrophage Colony-Stimulating Factor; Intercellular Adhesion Molecule-1; Macrophages, Peritoneal; Male; Mice; Mice, Inbred ICR; Peritoneal Neoplasms; Receptors, CXCR4; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A

Dendritic cells regulate immune responses to microbial products and play a key role in ulcerative colitis (UC) pathology. We determined the immunomodulatory effects of probiotic strain Lactobacillus casei Shirota (LcS) on human DC from healthy controls and active UC patients.. Human blood DC from healthy controls (control-DC) and UC patients (UC-DC) were conditioned with heat-killed LcS and used to stimulate allogeneic T cells in a 5-day mixed leucocyte reaction.. UC-DC displayed a reduced stimulatory capacity for T cells (P < 0.05) and enhanced expression of skin-homing markers CLA and CCR4 on stimulated T cells (P < 0.05) that were negative for gut-homing marker β7. LcS treatment restored the stimulatory capacity of UC-DC, reflecting that of control-DC. LcS treatment conditioned control-DC to induce CLA on T cells in conjunction with β7, generating a multihoming profile, but had no effects on UC-DC. Finally, LcS treatment enhanced DC ability to induce TGFβ production by T cells in controls but not UC patients.. We demonstrate a systemic, dysregulated DC function in UC that may account for the propensity of UC patients to develop cutaneous manifestations. LcS has multifunctional immunoregulatory activities depending on the inflammatory state; therapeutic effects reported in UC may be due to promotion of homeostasis.

Topics: Cell Proliferation; Cells, Cultured; Colitis, Ulcerative; Cytokines; Dendritic Cells; Flow Cytometry; Homeostasis; Humans; Inflammation; Lacticaseibacillus casei; Lymphocyte Activation; Probiotics; T-Lymphocytes; Transforming Growth Factor beta

In this study, we examined the therapeutic effects of an immune-stimulating peptide, WKYMVm, in ulcerative colitis. The administration of WKYMVm to dextran sodium sulfate (DSS)-treated mice reversed decreases in body weight, bleeding score and stool score in addition to reversing DSS-induced mucosa destruction and shortened colon. The WKYMVm-induced therapeutic effect against ulcerative colitis was strongly inhibited by a formyl peptide receptor (FPR) 2 antagonist, WRWWWW, indicating the crucial role of FPR2 in this effect. Mechanistically, WKYMVm effectively decreases intestinal permeability by stimulating colon epithelial cell proliferation. WKYMVm also strongly decreases interleukin-23 and transforming growth factor-β production in the colon of DSS-treated mice. We suggest that the potent immune-modulating peptide WKYMVm and its receptor FPR2 may be useful in the development of efficient therapeutic agents against chronic intestinal inflammatory diseases.

Topics: Adjuvants, Immunologic; Animals; Caco-2 Cells; Cell Proliferation; Colitis, Ulcerative; Colon; Humans; Interleukin-23; Intestinal Mucosa; Mice; Mice, Inbred C57BL; Oligopeptides; Permeability; Receptors, Formyl Peptide; Transforming Growth Factor beta

The 6 types of cross-linked autobodies of one family where identified during relapsing course of Ulcerativ Colities (UC) acompanied with deterioration of clinical and endoscopic activity and increasing rate of acut inflammatory phase (CRP, number of leukocytes and erythrocyte sedimentation rate) of the disease. On the background of transplantation of mesenchymal bone marrow stromal cells (BM MSC), and despit the identification of six types of autoantibodies to antigens of neutrophils, was observed moderate activity of UC and low concentration of autoantibodies than in immunosuppressive therapy without BM MSC transplantation. Discovered anti-inflammatory effect of BM MSCs transplantation in UC may be explained by the systemic influence of immunosuppressive effect: it is known that the BM MSCs inhibit dendritic cells, T-and B-lymphocytes participating in the immune response, activate regulatory T-cells, which produce antinflammatory cytokines, IL-10 and TGF-1beta, which suppress the inflammatory process.

Topics: Adult; Aged; Allografts; Antibodies, Antinuclear; B-Lymphocytes; Colitis, Ulcerative; Cross Reactions; Dendritic Cells; Female; Humans; Interleukin-10; Male; Mesenchymal Stem Cell Transplantation; Middle Aged; T-Lymphocytes; Transforming Growth Factor beta

Epithelial to mesenchymal transition (EMT) seems to play an important role in the pathogenesis of fistulae, a common clinical complication of Crohn's disease (CD). TGFβ and interleukin-13 (IL-13) have been correlated with the onset of EMT-associated organ fibrosis and high levels of TGFβ have been shown in transitional cells (TCs) lining CD fistula tracts. This study investigated whether IL-13 could be involved in the pathogenesis of CD-associated fistulae.. Protein or mRNA levels in HT29 intestinal epithelial cells (IECs) or colonic lamina propria fibroblasts (CLPFs) were studied by western blotting or real-time PCR. CLPFs were isolated from non-inflammatory disease controls or patients with CD with or without fistulae and IL-13 levels were analysed in surgically removed fistula specimens by immunohistochemistry.. TGFβ induced IL-13 secretion in CLPFs from patients with fistulising CD. In fistula specimens high levels of IL-13 were detected in TCs covering fistula tracts. In HT29 IEC monolayers, IL-13 induced SLUG and β6-integrin mRNA, which are associated with cell invasion. HT29 spheroids completely disintegrated when treated with TGFβ for 7 days, whereas IL-13-treated spheroids did not show morphological changes. Here, TGFβ induced mRNA expression of SNAIL1 and IL-13, whereas IL-13 elevated SLUG and β6-integrin mRNA. An anti-IL-13 antibody was able to prevent IL-13-induced SLUG expression in HT29 IECs.. TGFβ induces IL-13 expression and an EMT-like phenotype of IECs, while IL-13 promotes the expression of genes associated with cell invasion. These findings suggest that TGFβ and IL-13 play a synergistic role in the pathogenesis of fistulae and inhibition of IL-13 might represent a novel therapeutic approach for fistula treatment.

Topics: Adult; Biomarkers; Blotting, Western; Case-Control Studies; Colitis, Ulcerative; Crohn Disease; Female; Fibroblasts; HT29 Cells; Humans; Integrin beta Chains; Interleukin-13; Intestinal Fistula; Intestinal Mucosa; Male; Middle Aged; Real-Time Polymerase Chain Reaction; Snail Family Transcription Factors; Transcription Factors; Transforming Growth Factor beta

Tissue inhibitor of metalloproteinases (TIMP)-3 is an inhibitor of matrix metalloproteinases, which regulates tissue inflammation, damage, and repair. We investigated the role of TIMP-3 in intestinal inflammation in human beings and mice.. We used real-time polymerase chain reaction and flow cytometry to measure levels of TIMP-3 in intestine samples from patients with Crohn's disease (CD) and those without (controls). We also analyzed TIMP-3 levels in lamina propria mononuclear cells (LPMCs) collected from biopsy samples of individuals with or without CD (controls) and then stimulated with transforming growth factor (TGF)-β1, as well as in biopsy samples collected from patients with CD and then incubated with a Smad7 anti-sense oligonucleotide (knock down). LPMCs and biopsy samples from patients with CD were cultured with exogenous TIMP-3 and levels of inflammatory cytokines were measured. We evaluated the susceptibility of wild-type, TIMP-3-knockout (TIMP-3-KO), and transgenic (TIMP-3-Tg) mice to induction of colitis with 2, 4, 6-trinitrobenzene-sulfonic-acid (TNBS), and the course of colitis in recombinase-activating gene-1-null mice after transfer of wild-type or TIMP-3-KO T cells.. Levels of TIMP-3 were reduced in intestine samples from patients with CD compared with controls. Incubation of control LPMCs with TGF-β1 up-regulated TIMP-3; knockdown of Smad7, an inhibitor of TGF-β1, in biopsy samples from patients with CD increased levels of TIMP-3. Exogenous TIMP-3 reduced levels of inflammatory cytokines in CD LPMCs and biopsy samples. TIMP-3-KO mice developed severe colitis after administration of TNBS, whereas TIMP-3-Tg mice were resistant to TNBS-induced colitis. Reconstitution of recombinase-activating gene-1-null mice with T cells from TIMP-3-KO mice increased the severity of colitis, compared with reconstitution with wild-type T cells.. TIMP-3 is down-regulated in inflamed intestine of patients with CD. Its expression is regulated by TGF-β1, and knock-down of Smad7 in intestinal tissues from patient with CD up-regulates TIMP-3. Loss or reduction of TIMP-3 in mice promotes development of colitis.

Topics: Adult; Aged; Amyloid Precursor Protein Secretases; Animals; Cells, Cultured; Colitis; Colitis, Ulcerative; Crohn Disease; Cytokines; Down-Regulation; Gene Knockdown Techniques; Gene Knockout Techniques; Humans; Intestinal Mucosa; Leukocytes, Mononuclear; Mice; Mice, Knockout; Mice, Transgenic; Middle Aged; Oligonucleotides, Antisense; RNA, Messenger; Smad7 Protein; Tissue Inhibitor of Metalloproteinase-3; Transforming Growth Factor beta; Trinitrobenzenesulfonic Acid

Both chronic inflammation and somatic mutations likely contribute to the pathogenesis of ulcerative colitis (UC)-associated dysplasia and cancer. On the other hand, both tumor suppression and oncogenesis can result from transforming growth factor (TGF)-β signaling. TGF-β type I receptor (TβRI) and Ras-associated kinases differentially phosphorylate a mediator, Smad3, to become C-terminally phosphorylated Smad3 (pSmad3C), linker phosphorylated Smad3 (pSmad3L), and both C-terminally and linker phosphorylated Smad3 (pSmad3L/C). The pSmad3C/p21(WAF1) pathway transmits a cytostatic TGF-β signal, while pSmad3L and pSmad3L/C promote cell proliferation by upregulating c-Myc oncoprotein. The purpose of this study was to clarify the alteration of Smad3 signaling during UC-associated carcinogenesis.. By immunostaining and immunofluorescence, we compared pSmad3C-, pSmad3L-, and pSmad3L/C-mediated signaling in colorectal specimens representing colitis, dysplasia, or cancer from eight UC patients with signaling in normal colonic crypts. We also investigated p53 expression and mutations of p53 and K-ras genes. We further sought functional meaning of the phosphorylated Smad3-mediated signaling in vitro.. As enterocytes in normal crypts migrated upward toward the lumen, cytostatic pSmad3C/p21(WAF1) tended to increase, while pSmad3L/c-Myc shown by progenitor cells gradually decreased. Colitis specimens showed prominence of pSmad3L/C/c-Myc, mediated by TGF-β and tumor necrosis factor (TNF)-α, in all enterocyte nuclei throughout entire crypts. In proportion with increases in frequency of p53 and K-ras mutations during progression from dysplasia to cancer, the oncogenic pSmad3L/c-Myc pathway came to be dominant with suppression of the pSmad3C/p21(WAF1) pathway.. Oncogenic Smad3 signaling, altered by chronic inflammation and eventually somatic mutations, promotes UC-associated neoplastic progression by upregulating growth-related protein.

Topics: Adenocarcinoma; Adult; Blotting, Western; Cell Transformation, Neoplastic; Chronic Disease; Colitis, Ulcerative; Colorectal Neoplasms; Disease Progression; Female; Fluorescent Antibody Technique; Humans; Immunoenzyme Techniques; Immunoprecipitation; Male; Middle Aged; Phosphorylation; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; Protein Serine-Threonine Kinases; Receptor, Transforming Growth Factor-beta Type I; Receptors, Transforming Growth Factor beta; Signal Transduction; Smad3 Protein; Transforming Growth Factor beta

To investigate the effects of human gut micro-organisms on cytokine production by human intestinal cell lines.. Quantitative real-time PCR assays were developed to measure the production of pro-inflammatory (IL-1α, IL-6, IL-18 and TNFα) and anti-inflammatory (TGF-β1, TGF-β2, TGF-β3, IL-4 and IL-10) cytokines in HT-29 and Caco-2 cell lines. They were co-cultured with a range of mucosal bacteria isolated from ulcerative colitis patients, together with lactobacilli and bifidobacteria obtained from healthy people. HT-29 cells were also co-cultured with Campylobacter jejuni, enterotoxigenic Escherichia coli (ETEC), enteropathogenic E. coli and Salmonella typhimurium. The majority of commensal bacteria tested suppressed the expression of anti-inflammatory cytokine mRNA, increased IL-18, reduced IL-1α, and with the exception of nonpathogenic E. coli, reduced TNF-α. All overtly pathogenic species increased both pro-inflammatory and anti-inflammatory cytokine mRNA..   Commensal and pathogenic species induced fundamentally different cytokine responses in human intestinal epithelial cell lines.. Interactions between commensal bacteria tested in this study and the innate immune system were shown to be anti-inflammatory in nature, in contrast to the pathogenic organisms investigated. These data contribute towards our understanding of how potential probiotic species can be used to suppress the pro-inflammatory response in inflammatory bowel disease.

Topics: Bacterial Physiological Phenomena; Bifidobacterium; Caco-2 Cells; Campylobacter jejuni; Coculture Techniques; Colitis, Ulcerative; Cytokines; Epithelial Cells; Escherichia coli; HT29 Cells; Humans; Interleukins; Intestinal Mucosa; Lactobacillus; Probiotics; Salmonella typhimurium; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Mϕs promote tissue injury or repair depending on their activation status and the local cytokine milieu. It remains unclear whether the immunosuppressive effects of transforming growth factor β (TGF-β) serve a nonredundant role in Mϕ function in vivo. We generated Mϕ-specific transgenic mice that express a truncated TGF-β receptor II under control of the CD68 promoter (CD68TGF-βDNRII) and subjected these mice to the dextran sodium sulfate (DSS) model of colitis. CD68TGF-βDNRII mice have an impaired ability to resolve colitic inflammation as demonstrated by increased lethality, granulocytic inflammation, and delayed goblet cell regeneration compared with transgene negative littermates. CD68TGF-βDNRII mice produce significantly less IL-10, but have increased levels of IgE and numbers of IL-33+ Mϕs than controls. These data are consistent with associations between ulcerative colitis and increased IL-33 production in humans and suggest that TGF-β may promote the suppression of intestinal inflammation, at least in part, through direct effects on Mϕ function.

Topics: Animals; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Blotting, Southern; Colitis; Colitis, Ulcerative; Dextran Sulfate; Disease Models, Animal; Flow Cytometry; Immunoglobulin E; Interleukin-33; Interleukins; Macrophages; Mice; Mice, Transgenic; Promoter Regions, Genetic; Receptors, Transforming Growth Factor beta; Transforming Growth Factor beta

The intestinal immune system is constantly challenged by foreign antigens and commensal bacteria. Therefore, proper control of the intestinal microenvironment is required. One important arm of this regulatory network consists of regulatory T cells. In contrast to CD4(+) Foxp3(+) regulatory T cells, which have been well characterized, immunomodulatory CD8(+) T cells that express Foxp3 are less well defined in terms of their generation and function. Failures of these regulatory mechanisms contribute to the development of inflammatory bowel disease. In this study we demonstrate that the frequency of CD8(+) Foxp3(+) T cells is reduced in the peripheral blood of patients with ulcerative colitis. As these cells might play a currently underestimated role in the maintenance of intestinal homeostasis, we have investigated human and murine CD8(+) Foxp3(+) T cells generated by stimulating naive CD8(+) T cells in the presence of transforming growth factor-β and retinoic acid, mediators that are abundantly produced in the intestinal mucosa. These CD8(+) Foxp3(+) fully competent regulatory T cells show strong expression of regulatory molecules CD25, Gpr83 and CTLA-4 and exhibit cell-cell contact-dependent immunosuppressive activity in vitro. Our study illustrates a previously unappreciated critical role of CD8(+) Foxp3(+) T cells in controlling potentially dangerous T cells and in the maintenance of intestinal homeostasis.

Topics: Adult; Aged; Aged, 80 and over; Animals; Antigens, CD; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cell Communication; Cell Count; Cell Proliferation; Coculture Techniques; Colitis, Ulcerative; CTLA-4 Antigen; Cytotoxicity, Immunologic; Dendritic Cells; Female; Forkhead Transcription Factors; Gene Expression; Granzymes; Green Fluorescent Proteins; Humans; Immune Tolerance; Immunophenotyping; Interferon-gamma; Interleukin-2 Receptor alpha Subunit; Leukocytes, Mononuclear; Lymphocyte Activation; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Mice, Transgenic; Middle Aged; Pore Forming Cytotoxic Proteins; T-Lymphocyte Subsets; Transforming Growth Factor beta; Tretinoin

Saccharomyces boulardii (Sb) is a probiotic yeast that has demonstrated efficacy in pilot studies in patients with inflammatory bowel disease (IBD). Microbial antigen handling by dendritic cells (DC) is believed to be of critical importance for immunity and tolerance in IBD. The aim was to characterize the effects of Sb on DC from IBD patients. Highly purified (>95%), lipopolysaccharide-stimulated CD1c(+)CD11c(+)CD123(-) myeloid DC (mDC) from patients with ulcerative colitis (UC; n = 36), Crohn's disease (CD; n = 26), or infectious controls (IC; n = 4) were cultured in the presence or absence of fungal supernatant from Sb (SbS). Phenotype and cytokine production and/or secretion of IBD mDC were measured by flow cytometry and cytometric bead arrays, respectively. T cell phenotype and proliferation were assessed in a mixed lymphocyte reaction (MLR) with allogenic CD4(+)CD45RA(+) naïve T cells from healthy donors. Mucosal healing was investigated in epithelial wounding and migration assays with IEC-6 cells. SbS significantly decreased the frequency of CD40-, CD80-, and CD197 (CCR7; chemokine receptor-7)-expressing IBD mDC and reduced their secretion of tumor necrosis factor (TNF)-α and interleukin (IL)-6 while increasing IL-8. In the MLR, SbS significantly inhibited T cell proliferation induced by IBD mDC. Moreover, SbS inhibited T(H)1 (TNF-α and interferon-γ) polarization induced by UC mDC and promoted IL-8 and transforming growth factor-β-dependent mucosal healing. In summary, we provide novel evidence of synergistic mechanisms how Sb controls inflammation (inhibition of T cell costimulation and inflammation-associated migration and mobilization of DC) and promotes epithelial restitution relevant in IBD.

Topics: B7-1 Antigen; CD40 Antigens; Cell Division; Cell Movement; Cells, Cultured; Colitis, Ulcerative; Crohn Disease; Dendritic Cells; Female; Humans; Immunotherapy; Interleukin-6; Interleukin-8; Lymphocyte Culture Test, Mixed; Male; Probiotics; Receptors, CCR7; Saccharomyces; T-Lymphocytes; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Gene expression of transforming growth factor-beta (TGF-beta) is needed to induce expression of transcription factor forkhead box P3 (Foxp3), which is required for the development and function of regulatory T (Treg) cells. The number of circulating Treg cells and the level of Foxp3 expression increase during granulocyte and monocyte apheresis (GMA), a useful therapy for ulcerative colitis. However, the mechanism underlying GMA-induced Foxp3 expression is unknown. We found that the level of TGF-beta mRNA in peripheral blood mononuclear cells (PBMCs) was augmented just after treatment of peripheral blood with a GMA carrier, cellulose acetate beads, in vitro and that Foxp3 expression in PBMCs increased after culturing these cells for 5 days after the treatment. The augmentation of TGF-beta expression was observed in CD3(-) PBMCs but not in CD3(+) T cells. Furthermore, the increase in Foxp3 expression in T cells depended on co-culture with CD3(-) PBMCs. We conclude that cellulose acetate beads have an ability to induce Foxp3 expression in peripheral blood T cells via augmentation of TGF-beta expression in CD3(-) PBMCs.

Topics: Blood Component Removal; CD3 Complex; Cellulose; Coculture Techniques; Colitis, Ulcerative; Forkhead Transcription Factors; Granulocytes; Humans; Immunophenotyping; Microspheres; Monocytes; T-Lymphocytes, Regulatory; Transcriptional Activation; Transforming Growth Factor beta

Interleukin-33 (IL-33) is a novel member of the interleukin-1 family that induces mucosal pathology in vivo and may drive fibrosis development and angiogenesis. To address its potential role in inflammatory bowel disease, we explored its tissue expression in biopsy specimens from untreated ulcerative colitis patients, observing a 2.6-fold up-regulation of IL-33 mRNA levels, compared to controls. Immunohistochemical analyses of surgical specimens showed that a prominent source of IL-33 in ulcerative colitis lesions were ulceration-associated myofibroblasts that co-expressed the fibroblast marker heat shock protein 47, platelet-derived growth factor receptor (PDGFR)β, and, in part, the myofibroblast marker α-smooth muscle actin (SMA). In contrast, IL-33-positive myofibroblasts were almost absent near the deep fissures seen in Crohn's disease. A screen of known and putative activators of IL-33 in cultured fibroblasts revealed that the Toll-like receptor-3 agonist poly (I:C) was among the strongest inducers of IL-33 and that it synergized with transforming growth factor-β, a combination also known to boost myofibroblast differentiation. Experimental wound healing in rat skin revealed that the de novo induction of IL-33 in pericytes and the possible activation of scattered, tissue-resident IL-33(+)PDGFRβ(+)αSMA(-) fibroblast-like cells were early events that preceded the later appearance of IL-33(+)PDGFRβ(+)αSMA(+) cells. In conclusion, our data point to a novel role for IL-33 in mucosal healing and wound repair and to an interesting difference between ulcerative colitis and Crohn's disease.

Topics: Animals; Biopsy; Case-Control Studies; Cells, Cultured; Colitis, Ulcerative; Colon; Crohn Disease; Gene Expression; Humans; Inflammatory Bowel Diseases; Interleukin-33; Interleukins; Myofibroblasts; Poly I-C; Rats; Toll-Like Receptor 3; Transforming Growth Factor beta; Wound Healing

Ulcerative colitis (UC) and Crohn's disease (CD) are considered to be immunologically mediated disorders that share certain features with murine models of colitis. Whether any of these models are physiologically relevant to the human condition remains controversial. The hypothesis is that increased amounts of antibodies neutralizing transforming growth factor (TGF)-beta, interleukin (IL)-2 or IL-10 create a relative immunodeficient state in inflammatory bowel disease (IBD) that predisposes to disease. To evaluate this, serum samples from patients with UC or CD and from normal healthy individuals were studied by enzyme-linked immunosorbent assays. Antibodies recognizing TGF-beta were most prevalent in UC (P<0.01); anti-IL-10 antibodies were elevated in CD (P<0.05), while anti-IL-2 antibodies were the same for all three groups. Importantly, the percentage of IBD patients with at least one of the antibody levels greater than any control value was 30% for UC and 33% for CD. To verify the presence of these antibodies, immobilized TGF-beta was exposed to UC sera and the attached proteins identified by Western blot assay. The proteins proved to be exclusively immunoglobulin (Ig) G. To evaluate the neutralizing activity of these antibodies, cytokine-specific IgG from subjects in each group of patients was incubated with TGF-beta, IL-2 or IL-10 before addition to a bioassay with changes in viability determined by a colorimetric analysis. Antibodies from most individuals in all three groups neutralized the action of each cytokine. This study shows that about one-third of IBD patients may have a relative deficiency of TGF-beta, IL-2 or IL-10 due to an increase in neutralizing antibodies in their sera.

Topics: Adult; Analysis of Variance; Autoantibodies; Case-Control Studies; Cell Line, Tumor; Colitis, Ulcerative; Crohn Disease; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunoglobulin G; Interleukin-10; Interleukin-2; Male; Middle Aged; Statistics, Nonparametric; Transforming Growth Factor beta

Interleukin 17 (IL17) is now known to be involved in a number of chronic inflammatory disorders. However, the mechanisms regulating its production in inflammatory bowel disease (IBD) are still unclear.. Endoscopic biopsies or surgical specimens were taken from inflamed and uninflamed colonic mucosa of 72 patients with IBD (38 with Crohn's disease and 34 with ulcerative colitis), and normal colon of 38 control subjects. IL17 and interferon gamma (IFNgamma) were detected by ELISA in the supernatants of biopsies cultured ex vivo, and anti-CD3/CD28-stimulated lamina propria mononuclear cells (LPMCs) incubated with IL12, IL23, IL1beta plus IL6, transforming growth factor beta1 (TGFbeta1), or anti-IL21 neutralising antibody. Intracellular flow cytometry was performed to analyse mucosal Th17 and Th1/Th17 cells.. IL17 production by organ culture biopsies was higher in IBD inflamed mucosa than IBD uninflamed mucosa and controls, and was equivalent in amount to IFNgamma. Anti-CD3/CD28-stimulated IBD LPMCs produced higher IL17 amounts compared to controls. The percentages of Th17 and Th1/Th17 cells were increased in patients with IBD. IL23 and IL1beta plus IL6 had no effect on IBD LPMC production of IL17; however, IL12 markedly increased IFNgamma production and decreased IL17 production. TGFbeta1 dose-dependently decreased IFNgamma, but had no significant inhibitory effect on IL17 production. Blocking IL21 significantly downregulated IL17 production.. Our findings support a role for IL12, TGFbeta and IL21 in modulating IL17/IFNgamma production in IBD. The abundant IL17 in inflamed IBD mucosa may help explain the relative lack of efficacy of anti-IFNgamma antibodies in clinical trials of Crohn's disease.

Topics: Adolescent; Adult; Cells, Cultured; Colitis, Ulcerative; Crohn Disease; Cytokines; Dose-Response Relationship, Immunologic; Extracellular Matrix Proteins; Humans; Immunity, Mucosal; Inflammation Mediators; Inflammatory Bowel Diseases; Interferon-gamma; Interleukin-17; Intestinal Mucosa; Middle Aged; Organ Culture Techniques; T-Lymphocyte Subsets; Th1 Cells; Transforming Growth Factor beta; Young Adult

To investigate if the clinical efficacy of granulocytes and monocytes by adsorption (GMA) is associated with an increased frequency of peripheral regulatory T cells (Tregs), as these cells have proven to be successful in suppressing inflammatory bowel disease (IBD) in animal models.. We report four cases of corticosteroid-dependent ulcerative colitis (UC) and two Crohn's disease (CD) cases with severe cutaneous lesions who received GMA therapy. The frequency of CD4+ CD25(high) (Tregs) in peripheral blood was analyzed by flow cytometry and the expression of FoxP3 and TGF beta in purified CD4+ T cells was determined by real time PCR prior to and one month after the last apheresis session, and at the time of endoscopic and clinical assessing.. Increased expression of Fox P3 mRNA was found in all five patients who responded to cytapheresis with remission of clinical symptoms, mucosal inflammation and cutaneous lesions, and an increased frequency of circulating Tregs was found in four patients. These changes were not observed in the patient with UC who did no respond to GMA. Variations in TGF-beta (mRNA) did not parallel that of FoxP3 mRNA.. The clinical efficacy of GMA on IBD and related extra intestinal manifestations was associated with an expansion of circulating CD4+ CD25+ Tregs and higher expression of FoxP3 in CD4+ T cells. Accordingly, an elevated CD4+ CD25+ FoxP3 may be a valuable index of remission in patients with IBD and other chronic relapsing-remitting inflammatory conditions during treatment with GMA.

Topics: Adult; CD4 Antigens; Colitis, Ulcerative; Crohn Disease; Cytapheresis; Female; Forkhead Transcription Factors; Humans; Interleukin-2 Receptor alpha Subunit; Male; RNA, Messenger; Skin Diseases; T-Lymphocytes, Regulatory; Transforming Growth Factor beta; Treatment Outcome

Cytokines and growth factors play a major role in the dysregulated immune response in inflammatory bowel disease (IBD). We hypothesized that significant differences exist between the serum cytokine and growth factor profiles of pediatric IBD patients with active disease (AD) and those in remission, and that levels of some of these soluble mediators may be used to define regulators in IBD and determine disease activity.. Eighty-eight consecutive patients with confirmed Crohn's disease (CD) and ulcerative colitis (UC) seen at the Duke Children's Hospital were prospectively enrolled and a serum sample was obtained. Data were recorded at the time of serum collection to calculate disease activity indices. The relative expression of 78 cytokines, growth factors, and soluble receptors was determined using proprietary antibody-based protein microarrays amplified by rolling circle amplification. SPSS 8 (SPSS Inc., Chicago, IL) was used to compare protein profiles for CD and UC patients in clinical remission (CR) versus AD.. Sixty-five CD patients and 23 UC patients were enrolled. Forty-one CD patients had available samples and PCDAI results. Twenty-two patients were in remission PCDAI < or = 12.5 (median 5), 19 patients had disease activity >15 (median 30). Univariate analysis revealed that PLGF, IL-7, IL-12p40, and TGF-beta1 cytokine levels were significantly elevated for patients in CR versus AD (p < 0.01). Twelve UC serum samples had Seo/Truelove Witt AI for analysis. Five patients were in remission by TW AI and Seo AI < or =110 and 7 patients had active mild-to-severe disease by TW and Seo AI >110. Only one cytokine, IL12p40, showed significance between CR versus AD (p < 0.02).. Surprisingly, we found no differences in circulating levels of proinflammatory cytokines but found that pediatric IBD patients in remission compared to those with AD had higher levels of specific circulating cytokines, including the regulatory cytokines IL-12p40 and TGF-beta1. It may be that these cytokines directly regulate intestinal inflammation in IBD or reflect the activity of T regulatory cells in negatively regulating the inflammatory response. Further studies will be needed to validate our results to define the molecular pathways involved in the intestinal immune response in man.

Topics: Adolescent; Child; Child, Preschool; Colitis, Ulcerative; Crohn Disease; Cytokines; Female; Growth Substances; Humans; Infant; Inflammation Mediators; Interleukin-12; Interleukin-12 Subunit p40; Interleukin-7; Male; Placenta Growth Factor; Pregnancy Proteins; Protein Array Analysis; Protein Subunits; Transforming Growth Factor beta; Transforming Growth Factor beta1

To elucidate the differences in expression of TGFbeta1 and TGFbeta1 mRNA between ulcerative colitis (UC), infectious colitis (IC) and normal control.. TGFbeta1 in colonic mucosa was detected by immunohistochemistry (IHC). TGFbeta1 mRNA was detected by hybridization in situ.. There was no difference in detecting TGFbeta1 expression and TGFbeta1 mRNA expression in colonic mucosa between UC group and IC group (P>0.05), but the expression rates for the two groups were significantly higher than those for normal control (P<0.001). The expression of TGFbeta1 in colonic mucosa of UC group was noted to have a positive correlation with UC histological grade (r=0.462, P=0.002).. Enhanced TGFbeta1 production in the colonic mucosa of UC patients can not inhibit proinflammatory cytokine production and hence can not get control of inflammation; this finding suggests the possible presence of TGFbeta1 signaling defects in the cases of UC. TGFbeta1 may serve as a disease activity marker of ulcerative colitis.

Topics: Adult; Aged; Biomarkers; Colitis, Ulcerative; Colon; Female; Humans; Intestinal Mucosa; Male; Middle Aged; Transforming Growth Factor beta; Transforming Growth Factor beta1

Osteoporosis is a significant complication of a multifactoral etiology associated with inflammatory bowel disease. The aim of study was to evaluate the relationships between bone mineral density as well as bone turnover markers and inflammatory activity modulators (i.e., PGE2 and TGFbeta1) in ulcerative colitis (UC). Twenty-one active ulcerative colitis subjects and 14 healthy individuals were included into the study. We observed no significant differences in serum concentrations of osteoprotegerin and osteocalcin, as well as bone mineral density between UC patients and healthy individuals. Plasma concentrations of PGE2, TGFbeta1 and TNF-alpha were significantly higher in UC patients than in controls. Serum osteocalcin demonstrated a positive correlation with both serum PGE2 and plasma TGFbeta1. Moreover there was significant correlation between osteoprotegerin and TGFbeta1 as well as serum TNF-alpha concentrations. In conclusion a positive association between PGE2 and TGFbeta1 and bone formation markers-osteoprotegerin and osteocalcin, as well as a comparable BMD in UC patients and healthy individuals was shown. Our results may indicate that increase of PGE2 as well as TGFbeta1 concentrations may play a protective role against bone loss in ulcerative colitis patients.

Topics: Absorptiometry, Photon; Adult; Bone Density; Bone Remodeling; Colitis, Ulcerative; Dinoprostone; Female; Glycoproteins; Humans; Magnetic Resonance Imaging; Male; Osteocalcin; Osteoprotegerin; Receptors, Cytoplasmic and Nuclear; Receptors, Tumor Necrosis Factor; Transforming Growth Factor beta; Transforming Growth Factor beta1; Tumor Necrosis Factor-alpha

Studies aiming to define key cytokines in inflammatory bowel disease have been restricted to gene expression or protein quantitation but lack functional information on cytokine interactions. Some of the major cytokines that govern the extent and duration of the inflammatory process in ulcerative colitis (UC), appear to be interleukin 1 (IL-1), its natural inhibitor IL-1 receptor antagonist (IL-1ra) and transforming growth factor beta1 (TGF-beta 1). Indeed, as a predictor of inflammation, the mucosal status of IL-1, depicted as a ratio of IL-1ra/IL-1, has often been used.. Using an IL-1 bioassay and specific anti-cytokine antibodies we have identified the functional role of these cytokines and their interactions in mucosal biopsy samples taken from patients with UC.. Compared with control specimens, the secreted and tissue levels of IL-1 were consistently raised in UC samples. Levels of IL-1, rather than IL-1ra or the ratio of IL-1ra/IL-1, most closely mirrored the severity of inflammation. Using specific antibodies we showed that IL-1ra and TGF-beta 1 appear to modulate the degree of inflammation at different stages of the inflammatory process. Only in severely inflamed tissue, when IL-1 levels were high did IL-1ra inhibit IL-1-induced activity. In contrast, the levels of TGF-beta 1, and its effect in controlling inflammation, was most marked in mild but not severe UC.. The functional roles of these cytokines in the inflammatory process can now be more carefully elucidated using a bioassay and specific neutralising antibodies.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Biopsy; Colitis, Ulcerative; Colon; Female; Humans; Interleukin 1 Receptor Antagonist Protein; Interleukin-1; Intestinal Mucosa; Male; Middle Aged; Sialoglycoproteins; Transforming Growth Factor beta; Transforming Growth Factor beta1

Intestinal fibrosis and strictures frequently occur in Crohn's disease but not ulcerative colitis. We have recently shown that, compared to myofibroblasts obtained from normal and ulcerative colitis tissue, myofibroblasts isolated from fibrotic Crohn's disease mucosal samples express significantly lower amounts of transforming growth factor (TGF)-beta 3, but the expression of TGF-beta 2 was significantly greater. We now report that in myofibroblast cultures established from fibrotic Crohn's disease mucosal samples there is significantly higher constitutive expression of tissue inhibitor of metalloproteinase (TIMP)-1 compared to similar cells isolated from normal or ulcerative colitis tissue. Myofibroblasts derived from normal mucosa and from mucosa affected by ulcerative colitis or Crohn's disease also expressed matrix metalloproteinase (MMP)-1, MMP-2, and MMP-3 but did not express MMP-9. Recombinant (r) TGF-beta 1 and rTGF-beta 2, but not rTGF-beta 3, induced expression of TIMP-1 in normal intestinal myofibroblasts. These studies illustrate a potential mechanism by which differential expression of isoforms of TGF-beta may lead to excessive deposition of extracellular matrix and stricture formation via TIMP-1-mediated inhibition of MMP activity.

Topics: Cells, Cultured; Colitis, Ulcerative; Crohn Disease; Fibroblasts; Gene Expression Regulation; Humans; Intestinal Mucosa; Matrix Metalloproteinase 1; Matrix Metalloproteinase 2; Matrix Metalloproteinase 3; Matrix Metalloproteinase 9; Recombinant Proteins; RNA, Messenger; Tissue Inhibitor of Metalloproteinase-1; Transcription, Genetic; Transforming Growth Factor beta

The aim of this study was to evaluate the effect of active ulcerative colitis (UC) treatment on transforming growth factor beta(1) (TGF-beta(1)) concentration in plasma and rectal mucosa measured in 28 patients. The highest plasma values were observed in patients with the severe course of the disease (74.2+/-14.0 ng/ml), and they were significantly higher than in the group with mild one (43.7+/-5.6 ng/ml). Mean TGF-beta(1) measured in mucosal samples from patients with severe UC (563+/-146 pg/mg protein) doubled values from patients with mild UC (286+/-65 pg/mg protein). Plasma and mucosal TGF-beta(1) correlated significantly with disease activity index (DAI) and clinical activity index (CAI). Plasma TGF-beta(1) correlated additionally with scored endoscopic degree of mucosal injury. Treatment caused significant decrease of plasma and mucosal TGF-beta(1) concentrations. Patients who responded completely had higher baseline plasma and mucosal TGF-beta(1) that decreased significantly after the treatment. These results show that plasma and mucosal concentrations of transforming growth factor beta(1) are strongly associated with ulcerative colitis activity, and successful treatment of the disease results with decrease of their levels. More effective response to the treatment can be achieved in patients with higher baseline concentrations of TGF-beta(1).

Topics: Administration, Oral; Adult; Aged; Colitis, Ulcerative; Female; Humans; Intestinal Mucosa; Male; Mesalamine; Middle Aged; Prednisone; Rectum; Severity of Illness Index; Sulfasalazine; Transforming Growth Factor beta; Transforming Growth Factor beta1

Transforming growth factor-beta (TGF-beta) is an inhibitory cytokine recognized as a key regulator of immunological homeostasis and inflammatory responses. TGF-beta is involved in experimental models of oral tolerance and in the pathogenesis of experimental colitis. Patients with inflammatory bowel disease (IBD) have inappropriate T cell responses to antigenic components of their own intestinal microflora, suggesting the presence of a disorder in the normal mucosal immune mechanism that ensures the down-regulation of responses to harmless constituents in the microflora. To evaluate the contribution of TGF-beta to this imbalance, we measured TGF-beta1 production by lamina propria mononuclear cells (LPMC) and T cells isolated from tissue specimens of patients with Crohn's disease (CD), ulcerative colitis (UC) and controls. Cells were cultured in the presence or absence of anti-CD2 plus anti-CD28 MoAbs and TGF-beta1 production in culture supernatants was measured by ELISA. LPMC isolated from CD patients produced significantly less TGF-beta1 than controls when stimulated via CD2 plus CD28 pathways (P = 0.001)] conversely, in UC patients increased production of TGF-beta1 compared to controls was observed (P = 0.0005). These differences were also observed with purified lamina propria (LP) T cells in both diseases and were associated with the presence of inflammation. Thus, TGF-beta1 production shows contrasting secretion in CD and in UC, probably as a consequence of the different Th polarization. The absolute or relative defect in TGF-beta1 production observed in CD and UC may contribute to the perpetuation of inflammation.

Topics: Adult; Aged; Cells, Cultured; Colitis, Ulcerative; Crohn Disease; Humans; Interferon-gamma; Interleukin-5; Leukocytes, Mononuclear; Middle Aged; Statistics, Nonparametric; T-Lymphocytes; Transforming Growth Factor beta

Ulcerative colitis (UC), a chronic inflammatory bowel disease, exhibits pronounced increase of T lymphocytes in the inflamed mucosa. To understand the role of intestinal T lymphocytes in the pathogenesis of UC their cytokine production in the mucosa was analysed. Intestinal T lymphocytes of UC, Crohn's disease and control patients were analysed for cytokine mRNA levels by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) directly after isolation without in vitro stimulation. Frequencies of cytokine positive cells were determined in UC and control colon by immunomorphometry. T lymphocytes in normal colon expressed interleukin (IL)-2, interferon (IFN)-gamma, tumour necrosis factor (TNF)-alpha and transforming growth factor (TGF)-beta1, but not IL-4, IL-5 or IL-10. In UC, a highly significant increase in IL-10 mRNA levels in T lymphocytes and an increased frequency of IL-10 positive cells was seen in colon. IL-10 mRNA levels were also elevated in T lymphocytes of the non-inflamed ileum and correlated with disease activity at both locations. CD4+ T lymphocytes were the major source of IL-10 mRNA. IL-2, IFN-gamma and TNF-alpha mRNA levels were decreased in colonic T lymphocytes, and virtually no IL-2, IFN-gamma, TNF-alpha or TGF-beta positive cells were detected in basal lymphoid aggregates. However, scattered IL-10 positive cells were found here. Lamina propria outside the aggregates contained IL-10-, IFN-gamma, TNF-alpha and TGF-beta but not IL-2 positive cells. T cells of UC patients did not express IL-4 or IL-5. Taken, together the data suggest a generalized activation of IL-10 producing CD4+ T cells along the intestine of UC patients. The local environment seems to determine the biological consequences of elevated IL-10.

Topics: Acute Disease; Adult; Aged; Biopsy; Case-Control Studies; CD4-Positive T-Lymphocytes; Colitis, Ulcerative; Colon; Female; Humans; Immunohistochemistry; Interferon-gamma; Interleukin-10; Interleukin-2; Interleukin-4; Interleukin-5; Intestinal Mucosa; Male; Middle Aged; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Patients with ulcerative colitis are at risk for colon cancer and frequently have microsatellite instability,which, in turn, is usually associated with inactivation of transforming growth factor (TGF) beta signaling. TGF-beta1 deficiency in mice can lead to colon cancer that is preceded by precancerous lesions having submucosal inflammation and hyperplastic crypts. Germ-free TGF-beta1-deficient mice are free of inflammation, hyperplasia, and cancer, but when reintroduced into a Helicobacter hepaticus-containing specific pathogen-free room, these lesions reappear. Because adenoma/carcinoma but not inflammation/hyperplasia is dependent on the genetic backgrounds tested, colitis is required, but not sufficient, for carcinogenesis. This animal model should provide insight into the protective role of TGF-beta1 in early stages of ulcerative colitis-associated human colon cancer.

Topics: Animals; Colitis, Ulcerative; Colonic Neoplasms; Disease Models, Animal; Female; Genetic Predisposition to Disease; Germ-Free Life; Humans; Male; Mice; Mice, Inbred C3H; Mice, Knockout; Transforming Growth Factor beta; Transforming Growth Factor beta1

Little information is available on the relative contribution of peptide growth factors and leukocyte-derived proteinases to the repair processes in inflammatory bowel disease (IBD). We investigated their possible roles in epithelial cell restitution and proliferation in patients with IBD.. The expression of hepatocyte growth factor (HGF), keratinocyte growth factor (KGF), transforming growth factor-beta (TGF-beta), and neutrophil elastase (NE) was examined in colonic mucosal tissues. The effects of organ culture supernatants of mucosal tissues on epithelial cell restitution and proliferation were analyzed in vitro using an intestinal cell line, IEC-6 cells.. Most organ cultures detected the presence of measurable levels of HGF, with a relative paucity of KGF and TGF-beta activity. Greater levels of HGF were obtained in the mucosa involved with IBD, especially in patients with ulcerative colitis (UC). The mucosa involved with UC also showed higher amounts of NE. The supernatants from the mucosa involved with UC possessed a prominent stimulatory effect on the restitution of IEC-6 cells. By contrast, significant suppression beyond baseline levels was observed for the proliferation of IEC-6 cells when they were incubated with recombinant HGF plus the supernatants from the mucosa involved with UC. This suppression was diminished considerably by preincubation of the supernatants with the anti-NE antibody.. HGF produced in the intestinal mucosa may be an important stimulator acting on epithelial cell restitution in patients with IBD. However, NE released in situ may impair mucosal repair through inhibiting epithelial cell proliferation in patients with UC.

Topics: Adult; Biomarkers; Cell Division; Cells, Cultured; Colitis, Ulcerative; Epithelial Cells; Female; Growth Substances; Hepatocyte Growth Factor; Humans; Immunohistochemistry; Intestinal Mucosa; Leukocyte Elastase; Male; Microscopy, Confocal; Middle Aged; Probability; Sampling Studies; Sensitivity and Specificity; Severity of Illness Index; Statistics, Nonparametric; Transforming Growth Factor beta

Intestinal strictures are frequent in Crohn's disease but not ulcerative colitis. We investigated the expression of transforming growth factor (TGF)-beta isoforms by isolated and cultured primary human intestinal myofibroblasts and the responsiveness of these cells and intestinal epithelial cells to TGF-beta isoforms. Normal intestinal myofibroblasts released predominantly TGF-beta(3) and ulcerative colitis myofibroblasts expressed both TGF-beta(1) and TGF-beta(3), whereas in myofibroblast cultures from fibrotic Crohn's disease tissue, there was significantly lower expression of TGF-beta(3) but enhanced release of TGF-beta(2). These distinctive patterns of TGF-beta isoform release were sustained through several myofibroblast passages. Proliferation of Crohn's disease myofibroblasts was significantly greater than that of myofibroblasts derived from normal and ulcerative colitis tissue. In contrast to cells from normal and ulcerative colitis tissue, neutralization of the three TGF-beta isoforms did not affect the proliferation of Crohn's disease intestinal myofibroblasts. Studies on the effect of recombinant TGF-beta isoforms on epithelial restitution and proliferation suggest that TGF-beta(2) may be the least effective of the three isoforms in intestinal wound repair. In conclusion, the enhanced release of TGF-beta(2) but reduced expression of TGF-beta(3) by Crohn's disease intestinal myofibroblasts, together with their enhanced proliferative capacity, may lead to the development of intestinal strictures.

Topics: Actins; Activin Receptors, Type I; Cell Division; Cells, Cultured; Colitis, Ulcerative; Colon; Crohn Disease; Desmin; Fibroblasts; Fibrosis; Gene Expression; Humans; Intestinal Mucosa; Isomerism; Microscopy, Electron; Protein Serine-Threonine Kinases; Receptor, Transforming Growth Factor-beta Type I; Receptor, Transforming Growth Factor-beta Type II; Receptors, Transforming Growth Factor beta; RNA, Messenger; Transforming Growth Factor beta; Transforming Growth Factor beta2; Transforming Growth Factor beta3; Vimentin; Wound Healing

Inflammatory bowel disease (IBD), the precise etiology of which remains unknown, is comprised of two forms of chronic intestinal inflammation; ulcerative colitis (UC) and Crohn's disease (CD). Recent evidence increasingly suggests that IBD is the result of dysfunctional immunoregulation manifested by inappropriate production of mucosal cytokines. An abnormal microcirculatory system has also been implicated in its pathogenesis. To elucidate the mechanism of ischemic change in IBD, we assesse serum concentration levels of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (b-FGF), and plasma level of endothelin-1 (ET-1). We also investigated the expression of VEGF, b-FGF, and transforming growth factor-beta1,2,3 (TGF-beta1,2,3) in tissue by immunostaining.. Blood samples were obtained from 11 patients with UC, 11 patients with CD, and 10 patients as controls. Paraffin-embedded samples were used for an immunohistochemical study.. The concentration levels (in picograms per milliliter) were as follows: for ET-1, UC: 127+/-47.0, CD: 167.3+/-35.1, and controls (asthma: 38.5+/-23.8, p < 0.01; diverticulitis: 40.5+/-25.6, p < 0.01), for b-FGF, UC: 9.2+/-1.9, CD: 9.1+/-1.5, and controls (asthma: 5.0+/-0, p < 0.01; diverticulitis: 5.0+/-0, p < 0.01), for VEGF, UC: 659.8+/-181.0, CD: 740.0+/-182.3, and controls (asthma: 193.7+/-58.7, p < 0.01; diverticulitis: 199.6+/-59.7, p < 0.01). The levels of VEGF and b-FGF were significantly higher in active IBD than those in the controls. There was a significant positive correlation among the serum levels of VEGF and b-FGF and the plasma level of ET-1; that is, elevated VEGF, b-FGF, and ET-1 levels correlated well with each other. Immunohistochemical studies showed increased venula in the submucosa and lamina propria. Overexpression of VEGF and b-FGF in endothelial cells was revealed and TGF-beta2 and TGF-beta3 were found in inflammatory cells of active IBD, but no change was observed around the vessels in the controls.. It is suggested that the reciprocal reaction of these cytokines may contribute to angiogenesis in IBD b inducing intestinal ischemia through vasoconstriction.

Topics: Adult; Colitis, Ulcerative; Colon; Crohn Disease; Endothelial Growth Factors; Female; Fibroblast Growth Factor 2; Humans; Ileum; Immunohistochemistry; Lymphokines; Male; Middle Aged; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

Topics: Adolescent; Adult; Aged; Case-Control Studies; Colitis, Ulcerative; Crohn Disease; Female; Humans; Male; Middle Aged; Polymorphism, Genetic; Polymorphism, Restriction Fragment Length; Polymorphism, Single Nucleotide; Transforming Growth Factor beta

Enhanced expression of transforming growth factor-beta1 (TGF-beta1) demonstrated in human colonic mucosa of patients with ulcerative colitis (UC), indicates its possible significance in the pathogenesis of this disease. The aim of this study was to evaluate plasma TGF-beta1 concentration in patients with different degrees of colonic mucosal injury, as a possible indicator of ulcerative colitis activity. TGF-beta1 concentration was measured with an enzyme immunoassay (EIA) in plasma of 45 patients with endoscopically confirmed UC. Values observed in UC patients (40.5+/-15.9 ng/ml) were significantly higher than in healthy people (18.3+/-11.6 ng/ml) and higher than in patients with irritable colon syndrome (ICS), (20.5+/-13.6 ng/ml). The highest plasma TGF-beta1 (58.6+/-112.1 ng/ml) was in patients with the severe UC course. TGF-beta1 level analysed in all UC patients revealed significant positive correlation with scored degree of mucosal injury (r=0.396;P<0.01). Among other possible laboratory markers of the disease activity, only C-reactive protein concentration demonstrated significant correlation. Enhanced production of TGF-beta1 can be related to inflammation activity. Measurement of plasma TGF-beta1 may be considered as a biomarker of the disease activity.

Topics: Adult; Aged; C-Reactive Protein; Case-Control Studies; Colitis, Ulcerative; Endoscopy; Female; Humans; Immunoenzyme Techniques; Male; Middle Aged; Transforming Growth Factor beta; Transforming Growth Factor beta1

TGF-beta1 functions as a negative regulator of T cell immune responses, signaling to target cells using the Smad family of proteins. We show here that Smad7, an inhibitor of TGF-beta1 signaling, is overexpressed in inflammatory bowel disease (IBD) mucosa and purified mucosal T cells. Both whole tissue and isolated cells exhibit defective signaling through this pathway, as measured by phospho-Smad3 immunoreactivity. Specific antisense oligonucleotides for Smad7 reduce Smad7 protein expression in cells isolated from patients with IBD, permitting the cells to respond to exogenous TGF-beta1. TGF-beta1 cannot inhibit proinflammatory cytokine production in isolated lamina propria mononuclear cells from patients with Crohn disease (CD), but inhibition of Smad7 restores TGF-beta1 signaling and enables TGF-beta1 to inhibit cytokine production. In inflamed mucosal tissue explants from patients with CD, inhibition of Smad7 also restores p-Smad3 and decreases proinflammatory cytokine production, an effect that is partially blocked by anti-TGF-beta1. These results show that Smad7 blockade of TGF-beta1 signaling helps maintain the chronic production of proinflammatory cytokines that drives the inflammatory process in IBD and that inhibition of Smad7 enables endogenous TGF-beta to downregulate this response.

Topics: Activin Receptors, Type I; Adolescent; Adult; Cells, Cultured; Child; Colitis, Ulcerative; Crohn Disease; Cytokines; DNA-Binding Proteins; Female; Gene Expression Regulation; Humans; Inflammatory Bowel Diseases; Interferon-gamma; Intestinal Mucosa; Male; Middle Aged; Oligodeoxyribonucleotides, Antisense; Organ Culture Techniques; Phosphorylation; Protein Processing, Post-Translational; Protein Serine-Threonine Kinases; Receptor, Transforming Growth Factor-beta Type I; Receptors, Transforming Growth Factor beta; Signal Transduction; Smad3 Protein; Smad7 Protein; Trans-Activators; Transforming Growth Factor beta; Transforming Growth Factor beta1; Tumor Necrosis Factor-alpha

An increased mucosal expression of transforming growth factor-beta (TGF-beta) and hepatocyte growth factor (HGF) has been reported in patients with active inflammatory bowel diseases (IBD) and in proximity to injured gastric and intestinal mucosal surfaces. The aim of this study was to measure systemic concentrations of TGF-beta and HGF and to assess their potential value to predict disease activity or severity of inflammation in patients with inflammatory bowel diseases.. Plasma HGF and TGF-beta1 peptide levels were determined in 29 patients with ulcerative colitis, 45 patients with Crohn's disease and 28 healthy controls using commercial ELISA assays. Peptide levels were correlated with disease activity indices and various laboratory parameters.. HGF and TGF-beta1 plasma levels were detected in all control and IBD subjects. Although a tendency towards increased HGF and TGF-beta1 peptide levels in IBD patients was observed, differences between groups were not significant In ulcerative colitis patients HGF plasma levels positively correlated with white blood cell counts and negatively correlated with serum albumin concentrations and haematocrit. In Crohn's disease patients, a positive correlation between TGF-beta and platelet count was observed.. HGF and TGF-beta1 plasma concentrations are not significantly different in IBD and healthy control subjects. Stratification of IBD patients according to disease activity did not reveal any substantial differences, suggesting that HGF and TGF-beta plasma levels have no value in the assessment of disease activity or severity of inflammation in patients with IBD.

Topics: Adult; Case-Control Studies; Colitis, Ulcerative; Crohn Disease; Enzyme-Linked Immunosorbent Assay; Female; Hepatocyte Growth Factor; Humans; Male; Severity of Illness Index; Transforming Growth Factor beta

To investigate serum levels of transforming growth factor-beta1 and interferon-gamma in active ulcerative colitis and to assess changes during treatment.. We prospectively evaluated serum from 25 patients with untreated active ulcerative colitis and 19 healthy controls. Disease activity score (DAI), serum transforming growth factor-beta1 and interferon-gamma levels were measured at baseline and after 7 days of conventional treatment. Disease activity score and transforming growth factor-beta1 were also assessed at 42 days.. Baseline transforming growth factor-beta1 levels were significantly higher in patients than in controls (P < 0.02). On the 7th day, transforming growth factor-beta1 levels increased only in patients who responded (P < 0. 01); variations in transforming growth factor-beta1 levels and disease activity score were inversely correlated (r=- 0.72, P < 0. 001). At day 42, serum transforming growth factor-beta1 decreased significantly compared with the 7th day (P < 0.05). While in controls, interferon-gamma was undetectable; untreated patients had higher, widely variable, levels. At day 7, responders had higher interferon-gamma values than unresponsive cases. Variations in interferon-gamma correlated moderately with changes in transforming growth factor-beta1 (r=0.53, P < 0.05). Cytokine response did not depend upon the type of treatment.. Both transforming growth factor-beta1 and interferon-gamma may play a role in the injury-repair process in active ulcerative colitis. Variations in circulating transforming growth factor-beta1 levels in the first week of treatment seem to be related to the therapeutic response.

Topics: Adult; Aged; Anti-Inflammatory Agents; Anti-Inflammatory Agents, Non-Steroidal; Case-Control Studies; Colitis, Ulcerative; Female; Humans; Interferon-gamma; Male; Middle Aged; Prednisone; Severity of Illness Index; Sulfasalazine; Transforming Growth Factor beta

Celiac disease is characterized by disturbed jejunal crypt-villus axis biology with immunoglobulin (Ig) A deposits underlining the epithelium. The aim of this study was to test whether celiac disease serum IgA (reticulin/endomysial autoantibodies) interferes with the mesenchymal-epithelial cell cross-talk.. Differentiation of T84 epithelial cells was induced with IMR-90 fibroblasts or transforming growth factor beta in three-dimensional collagen gel cultures. The effects of purified celiac IgA and monoclonal tissue transglutaminase antibodies (CUB7402) were studied by adding the antibodies to the cocultures.. Active celiac disease IgA, reactive for tissue transglutaminase, significantly inhibited T84 epithelial cell differentiation (P < 0.001) and increased epithelial cell proliferation (P = 0.024). Similar effects were obtained with antibodies against tissue transglutaminase.. Celiac disease-associated IgA class antibodies disturb transforming growth factor beta-mediated fibroblast-epithelial cell cross-talk in this in vitro crypt-villus axis model. This primary finding indicates that celiac disease-specific autoantibodies may also contribute to the formation of the gluten-triggered jejunal mucosal lesion in celiac disease.

Topics: Autoantibodies; Celiac Disease; Cell Differentiation; Cell Division; Cell Line; Coculture Techniques; Colitis, Ulcerative; Extracellular Matrix; Fibroblasts; GTP Phosphohydrolases; GTP-Binding Proteins; Humans; Immunoglobulin A; Intestinal Mucosa; Protein Glutamine gamma Glutamyltransferase 2; Reference Values; Transforming Growth Factor beta; Transglutaminases

Administration of insulin-like growth factor-I (IGF-I) results in selective growth of the gastrointestinal tract. We investigated the effects of IGF-I on the colonic damage induced by oral dextran sulphate sodium (DSS) in the rat.. Rats consumed 2% DSS in the drinking water for 10 days to induce colitis. Pumps were implanted on day 3 to deliver IGF-I for 7 days. Colonic histopathology and immunolocalization of transforming growth factor-beta1 (TGF-beta1) were assessed on day 10.. Compared with the colon of vehicle-treated rats consuming DSS, IGF-I increased the numbers of goblet cells by 76%, reduced the proportion of lamina propria cells expressing TGF-beta1, and reduced the thickness of submucosal and muscularis externa layers by 26% and 20%, respectively.. We conclude that the effects of IGF-I treatment on the colonic epithelium may be mediated directly, whereas the reduced inflammation in the mucosa and submucosa may be mediated by a mechanism other than up-regulation of TGF-beta1-mediated immunosuppression.

Topics: Animals; Body Weight; Colitis, Ulcerative; Colon; Dextran Sulfate; Dose-Response Relationship, Drug; Immunohistochemistry; Insulin-Like Growth Factor I; Intestinal Mucosa; Male; Organ Size; Rats; Rats, Sprague-Dawley; Transforming Growth Factor beta

Lipid peroxidation is a potential mechanism of bowel damage in colitis. The effect of oral consumption of a bovine whey-derived growth factor extract (WGFE) on lipid peroxidation was assessed using the ethane breath test in the dextran sulphate sodium (DSS) model of ulcerative colitis (UC) in rats.. Groups of rats consumed water (control), 2% DSS in drinking water, 2% DSS with a WGFE-supplemented diet, or 2% DSS plus prednisolone (1 mg x kg(-1) x day(-1)) for 6 weeks, changing to sulphasalazine (100 mg x kg(-1) x day(-1)) for the subsequent 4 weeks. Ethane breath tests were conducted on all animals on days 2, 4, 6, 8, and 10 (acute phase) and weeks 3, 6, and 9 (chronic phase) after commencement of DSS consumption.. There were no significant differences in ethane production between any groups during the acute phase. Ethane was significantly increased (P < 0.05) in rats consuming DSS alone in week 6 compared with control but had decreased to control levels by week 9. WGFE and conventional therapy were effective in suppressing ethane production in week 3.. WGFE is as effective as conventional therapies at limiting ethane production and thus ostensibly colonic lipid peroxidation in the early phases of experimental chronic UC.

Topics: Administration, Oral; Animals; Anti-Inflammatory Agents; Anti-Inflammatory Agents, Non-Steroidal; Breath Tests; Colitis, Ulcerative; Colon; Dextran Sulfate; Disease Models, Animal; Drug Therapy, Combination; Ethane; Growth Substances; Lipid Peroxidation; Male; Prednisolone; Rats; Rats, Sprague-Dawley; Sulfasalazine; Transforming Growth Factor beta

Transforming growth factors (TGFs) alpha and beta are key regulatory peptides that modulate mucosal cell populations critical to inflammatory bowel disease. The aim of this study was to assess TGF-alpha and TGF-beta expression in human colonic mucosa.. TGF-alpha and TGF-beta expression was assessed in colonic mucosa from patients with ulcerative colitis, patients with Crohn's disease, and controls by Northern blot analysis, in situ hybridization, and bioassay.. TGF-alpha messenger RNA expression localized to the villous tips of the small intestine and the surface epithelium of the colon. TGF-alpha expression was enhanced 2.3-fold in inactive ulcerative colitis mucosa relative to active ulcerative colitis, Crohn's disease, or normal controls. Enhanced expression correlated with duration of disease. TGF-beta expression was increased in affected mucosa from both patients with ulcerative colitis and Crohn's disease with active disease. TGF-beta1 messenger RNA expression in ulcerative colitis and Crohn's disease localized mostly to cells of the lamina propria with the highest concentration in inflammatory cells closest to the luminal surface.. TGF-alpha may contribute to epithelial hyperproliferation and the increased risk of malignancy in long-standing ulcerative colitis. TGF-beta may be a key cytokine during periods of active inflammation, modulating epithelial cell restitution and functional features of cells within the lamina propria.

Topics: Biological Assay; Blotting, Northern; Colitis, Ulcerative; Colon; Crohn Disease; Epithelium; Humans; In Situ Hybridization; Inflammatory Bowel Diseases; Intestinal Mucosa; RNA, Messenger; Transforming Growth Factor alpha; Transforming Growth Factor beta

This study was carried out to investigate the expression of various growth factors (GFs) involved in mucosal healing and thereby to clarify whether there are potential differences in the expression of GFs between normal colonic mucosa and the uninvolved mucosa of ulcerative colitis (UC). GF mRNA was investigated by reverse transcription polymerase chain reaction in colorectal biopsies from 20 normal controls and 15 UC patients. The positive rates (%) for mRNA expression for normal/UC were: epidermal growth factor (EGF) 65/53, transforming growth factor (TGF)-alpha 100/87, TGF-beta 1 60/33, insulin like growth factor-I 45/33, platelet-derived growth factor-A 55/67, basic fibroblast growth factor 0/0, hepatocyte growth factor (HGF) 50/53, EGF receptor 20/27, erb-B2 75/73, and HGF receptor (c-MET) 55/67. Semiquantitation of mRNA showed significantly lower expression of TGF-beta 1 (P < 0.05) in UC. Differences in expression and mRNA levels were not statistically significant for any other GFs. Our results indicate that mucosa in chronic persistent UC has a low basal expression of TGF-beta 1 mRNA, and, since TGF-beta 1 is a multifunctional GF that plays important roles in regulating repair and regeneration following tissue injury, this low expression may be partially responsible for the intractability of the disease.

Topics: Case-Control Studies; Colitis, Ulcerative; Colon; Female; Growth Substances; Humans; Intestinal Mucosa; Male; Middle Aged; Polymerase Chain Reaction; Receptors, Growth Factor; Rectum; RNA, Messenger; Transforming Growth Factor beta

Gastric cancers, sporadic colorectal cancers, and ulcerative colitis (UC)-associated colorectal carcinomas and dysplasias manifest microsatellite instability (MI); however, esophageal carcinomas rarely exhibit MI. Recently, a transforming growth factor-beta 1 type II receptor (TGF-beta 1RII) mutation in a coding microsatellite was described in primary colorectal carcinomas demonstrating MI. No previous studies of TGF-beta 1RII have addressed mechanisms of inactivation other than MI in human tumors; furthermore, MI-negative tumors have not been examined for TGF-beta 1RII mutation. We evaluated 138 primary human neoplasms for mutation in the poly-A microsatellite tract of TGF-beta 1RII. Additionally, a group of esophageal tumors was evaluated for the expression of TGF-beta 1RII messenger RNA (mRNA).. First, we determined whether MI was present at other chromosomal loci in these lesions. The poly-deoxyadenine (poly-A) microsatellite tract within the TGF-beta 1RII coding region was then PCR-amplified. In a group of MI-negative esophageal tumors, RT-PCR was performed to determine the expression of TGF-beta 1RII mRNA.. Among 17 MI+ UC specimens, 3 (18%) demonstrated TGF-beta 1RII poly-A tract mutation (2 cancers and 1 dysplasia), while 2 (4%) of 44 MI-negative UC specimens (1 dysplasia and 1 tumor), and 13 (81%) of 16 MI+ sporadic colorectal cancers, contained TGF-beta 1RII poly-A mutation. No gastric or esophageal tumors contained TGF-beta 1RII mutation. Among 21 MI-negative esophageal carcinomas. 6 cases (28.5%) had TGF-beta 1RII transcripts that were low or undetectable by RT-PCR.. Mutation within the poly-A microsatellite tract of TGF-beta 1RII occurs early in a subset of UC-neoplasms and commonly in sporadic colorectal cancers, but may be rare in MI+ gastric tumors. Diminished expression of TGF-beta 1RII mRNA in esophageal tumors suggests that mechanisms of inactivation in this gene other than MI play a role in esophageal carcinogenesis.

Topics: Colitis, Ulcerative; Colorectal Neoplasms; Esophageal Neoplasms; Humans; Microsatellite Repeats; Mutation; Protein Serine-Threonine Kinases; Receptor, Transforming Growth Factor-beta Type II; Receptors, Transforming Growth Factor beta; RNA, Messenger; Stomach Neoplasms; Transforming Growth Factor beta

Transforming growth factor (TGF)-beta inhibits cell proliferation and stimulates cell differentiation. For the signal transduction of TGF-beta, receptors I and II are required. The present study discloses the immunohistochemical localization of TGF-beta receptors I and II in inflammatory bowel disease. In the normal colon and small intestinal tissue, TGF-beta receptors were expressed in the epithelial cells in the upper crypts. In inflammatory bowel disease, a disordered expression pattern of receptors I and II was observed in the epithelial cells. Fibroblasts in the area of early stage fibrosis were positive for both receptors, while fibroblasts in area of advanced fibrosis lacked immunoreactivity for both receptors. Our study suggests that TGF-beta receptors I and II are important for the initial step of fibrosis.

Topics: Animals; Case-Control Studies; Colitis, Ulcerative; Colon; Crohn Disease; Fibroblasts; Humans; Ileum; Immunoenzyme Techniques; Intestinal Mucosa; Protein Serine-Threonine Kinases; Rabbits; Receptor, Transforming Growth Factor-beta Type II; Receptors, Transforming Growth Factor beta; Transforming Growth Factor beta