Differential regulation of interleukin-17 and interferon-y production in inflammatory bowel disease

PhDBackground and Aims. Interleukin (IL)-17 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. Methods. Endoscopic biopsies or surgical specimens were taken from inflamed and uninflamed colonic mucosa of 72 IBD patients (38 with Crohn’s disease and 34 with ulcerative colitis), and normal colon of 38 control subjects. IL-17 and interferon (IFN)- were detected by ELISA in the supernatants of biopsies cultured ex vivo, and anti-CD3/CD28-stimulated lamina propria mononuclear cells (LPMCs) incubated with IL-12, IL-23, IL-1β plus IL-6, transforming growth factor (TGF)-β1, or anti-IL-21 neutralising antibody. Intracellular flow cytometry was performed to analyse mucosal Th17 and Th1/Th17 cells. Results. IL-17 production by organ culture biopsies was higher in IBD inflamed mucosa than IBD uninflamed mucosa and controls, and was equivalent in amount to IFN-. Anti-CD3/CD28-stimulated IBD LPMCs produced higher IL-17 amounts compared to controls. The percentages of Th17 and Th1/Th17 cells were increased in IBD patients than controls. IL-23 and IL-1 plus IL-6 had no effect on IBD LPMC production of IL-17, however IL-12 markedly increased IFN- production and decreased IL-17 production. TGF-β1 dose-dependently decreased IFN-, but had no significant inhibitory effect on IL-17 production. Blocking IL-21 significantly down-regulated IL-17 production. Conclusions. Our findings support a role for IL-12, TGF- and IL-21 in modulating IL-17/IFN- production in IBD. The abundant IL-17 in inflamed IBD mucosa may help explain the relative lack of efficacy of anti-IFN- antibodies in clinical trials of Crohn’s disease

Proteolytic cleavage and loss of function of biologic agents that neutralize tumor necrosis factor in the mucosa of patients with inflammatory bowel disease

BACKGROUND & AIMS: Many patients with inflammatory bowel disease (IBD) fail to respond to anti–tumor necrosis factor (TNF) agents such as infliximab and adalimumab, and etanercept is not effective for treatment of Crohn’s disease. Activated matrix metalloproteinase 3 (MMP3) and MMP12, which are increased in inflamed mucosa of patients with IBD, have a wide range of substrates, including IgG1. TNFneutralizing agents act in inflamed tissues; we investigated the effects of MMP3, MMP12, and mucosal proteins from IBD patients on these drugs. METHODS: Biopsy specimens from inflamed colon of 8 patients with Crohn’s disease and 8 patients with ulcerative colitis, and from normal colon of 8 healthy individuals (controls), were analyzed histologically, or homogenized and proteins were extracted. We also analyzed sera from 29 patients with active Crohn’s disease and 33 patients with active ulcerative colitis who were candidates to receive infliximab treatment. Infliximab, adalimumab, and etanercept were incubated with mucosal homogenates from patients with IBD or activated recombinant human MMP3 or MMP12 and analyzed on immunoblots or in luciferase reporter assays designed to measure TNF activity. IgG cleaved by MMP3 or MMP12 and antihinge autoantibodies against neo-epitopes on cleaved IgG were measured in sera from IBD patients who subsequently responded (clinical remission and complete mucosal healing) or did not respond to infliximab. RESULTS: MMP3 and MMP12 cleaved infliximab, adalimumab, and etanercept, releasing a 32-kilodalton Fc monomer. After MMP degradation, infliximab and adalimumab functioned as F(ab’)2 fragments, whereas cleaved etanercept lost its ability to neutralize TNF. Proteins from the mucosa of patients with IBD reduced the integrity and function of infliximab, adalimumab, and etanercept. TNF-neutralizing function was restored after incubation of the drugs with MMP inhibitors. Serum levels of endogenous IgG cleaved by MMP3 and MMP12, and antihinge autoantibodies against neo-epitopes of cleaved IgG, were higher in patients who did not respond to treatment vs responders. CONCLUSIONS: Proteolytic degradation may contribute to the nonresponsiveness of patients with IBD to anti-TNF agents

New pathogenic paradigms in inflammatory bowel disease

Recent progresses in basic science have opened new pathogenic scenarios in inflammatory bowel disease. The T helper cell type (Th)1/Th2 paradigm has been outdated thanks to the advances in understanding the function of Th17 cells. Innate immunity, nonimmune cells, and defective tolerogenic mechanisms play a no less crucial role than do adaptive immunity, immune cells, and hyperactivation of effector mechanisms. These new paradigms, together with the no longer "static" but "dynamic" vision of intestinal inflammation, highlight new possible therapeutic targets in inflammatory bowel disease

Increased expression of mucosal addressin cell adhesion molecule-1 in the duodenum of patients with active celiac disease is associated with depletion of integrin α4β7-positive T cells in blood.

Mucosal addressin cell adhesion molecule 1, expressed on gut endothelial cells, in conjunction with integrin alpha(4)beta(7), expressed on lymphocytes, is critical in lymphocyte homing to the gut. The mucosal addressin cell adhesion molecule 1/integrin alpha(4)beta(7) pathway is involved in the pathogenesis of chronic intestinal inflammation by recruiting lymphocytes into inflamed gut. We explored the duodenal expression of mucosal addressin cell adhesion molecule 1 and the peripheral T-cell expression of integrin alpha(4)beta(7) in patients with celiac disease. Duodenal biopsies and a peripheral blood sample were collected from 15 celiac patients, before and after 12 months of gluten-free diet, and from 12 control subjects. Treated celiac biopsies were cultured with peptic-tryptic digest of gliadin and/or an anti-interferon alpha neutralizing antibody. Mucosal addressin cell adhesion molecule 1 was determined by confocal immunofluorescence microscopy and immunoblotting. Integrin beta(7)-positive T cells were analyzed by flow cytometry. Mucosal addressin cell adhesion molecule 1 expression was significantly higher in active celiac disease than in normal mucosa. After gluten-free diet, a dramatic reduction of mucosal addressin cell adhesion molecule 1 was also observed. No difference was seen between patients with celiac disease after treatment and controls. Ex vivo peptic-tryptic digest of gliadin challenge induced a marked increase of mucosal addressin cell adhesion molecule 1 expression. Blocking interferon alpha inhibited the peptic-tryptic digest of gliadin-induced mucosal addressin cell adhesion molecule 1 overexpression. The percentage of circulating beta(7)-positive T cells was significantly lower in untreated celiac disease in comparison to controls but normalized after gluten-free diet. Mucosal addressin cell adhesion molecule 1 is strongly up-regulated in active celiac disease dependent on interferon alpha and is associated with peripheral depletion of integrin alpha(4)beta(7)-expressing T cells. We conclude that mucosal addressin cell adhesion molecule 1 may represent an important determinant for the generation of mucosal damage in celiac disease

Recent advances in understanding ulcerative colitis

Ulcerative colitis, one of the two main forms of inflammatory bowel disease, is characterized by inflammation of the large bowel with constant involvement of the rectum, and a possible continuous retrograde distribution up to the cecum. Typical macroscopic lesions are mucosal ulcerations, with immune cell infiltration and cryptic abscesses at histology. Ulcerative colitis usually manifests with bloody diarrhea, is associated with a number of extra-intestinal manifestations, and may be acutely complicated by toxic megacolon. Longstanding disease may predispose to the development of colorectal cancer. Therapeutic options include mesalazine, corticosteroids, immunomodulators and biologic agents; however, if these treatments fail, the only available therapeutic choice remaining is the surgical removal of the colon. This review emphasizes novel concepts in the basic aspects of ulcerative colitis, and, in addition to the current clinical and diagnostic knowledge, it also describes new treatment options for this condition