Isolation of Mycobacterium avium Subspecies paratuberculosis Reactive CD4 T Cells from Intestinal Biopsies of Crohn’s Disease Patients

Background: Crohn’s disease (CD) is a chronic granulomatous inflammation of the intestine. The etiology is unknown, but an excessive immune response to bacteria in genetically susceptible individuals is probably involved. The response is characterized by a strong Th1/Th17 response, but the relative importance of the various bacteria is not known. Methodology/Principal Findings: In an attempt to address this issue, we made T-cell lines from intestinal biopsies of patients with CD (n = 11), ulcerative colitis (UC) (n = 13) and controls (n = 10). The T-cell lines were tested for responses to various bacteria. A majority of the CD patients with active disease had a dominant response to Mycobacterium avium subspecies paratuberculosis (MAP). The T cells from CD patients also showed higher proliferation in response to MAP compared to UC patients (p,0.025). MAP reactive CD4 T-cell clones (n = 28) were isolated from four CD patients. The T-cell clones produced IL-17 and/or IFN-c, while minimal amounts of IL-4 were detected. To further characterize the specificity, the responses to antigen preparations from different mycobacterial species were tested. One T-cell clone responded only to MAP and the very closely related M. avium subspecies avium (MAA) while another responded to MAP, MAA and Mycobacterium intracellulare. A more broadly reactive T-cell clone reacted to MAP1508 which belongs to the esx protein family. Conclusions/Significance: The presence of MAP reactive T cells with a Th1 or Th1/Th17 phenotype may suggest a possible role of mycobacteria in the inflammation seen in CD. The isolation of intestinal T cells followed by characterization of their specificity is a valuable tool to study the relative importance of different bacteria in CD

Diagnosis and management of adult coeliac disease: guidelines from the British Society of Gastroenterology

A multidisciplinary panel of 18 physicians and 3 non-physicians from eight countries (Sweden, UK, Argentina, Australia, Italy, Finland, Norway and the USA) reviewed the literature on diagnosis and management of adult coeliac disease (CD). This paper presents the recommendations of the British Society of Gastroenterology. Areas of controversies were explored through phone meetings and web surveys. Nine working groups examined the following areas of CD diagnosis and management: classification of CD; genetics and immunology; diagnostics; serology and endoscopy; follow-up; gluten-free diet; refractory CD and malignancies; quality of life; novel treatments; patient support; and screening for CD

A randomized trial of a transglutaminase 2 inhibitor for celiac disease

BACKGROUND In celiac disease, small intestinal transglutaminase 2 causes deamidation of glutamine residues in gluten peptides, which enhances stimulation of T cells and leads to mucosal injury. Inhibition of transglutaminase 2 is a potential treatment for celiac disease. METHODS In a proof-of-concept trial, we assessed the efficacy and safety of a 6-week treatment with ZED1227, a selective oral transglutaminase 2 inhibitor, at three dose levels as compared with placebo, in adults with well-controlled celiac disease who underwent a daily gluten challenge. The primary end point was the attenuation of gluten-induced mucosal damage, as measured by the ratio of villus height to crypt depth. Secondary end points included intraepithelial lymphocyte density, the Celiac Symptom Index score, and the Celiac Disease Questionnaire score (for assessment of health-related quality of life). RESULTS Of the 41 patients assigned to the 10-mg ZED1227 group, the 41 assigned to the 50-mg group, the 41 assigned to the 100-mg group, and the 40 assigned to the placebo group, 35, 39, 38, and 30 patients, respectively, had adequate duodenal-biopsy samples for the assessment of the primary end point. Treatment with ZED1227 at all three dose levels attenuated gluten-induced duodenal mucosal injury. The estimated difference from placebo in the change in the mean ratio of villus height to crypt depth from baseline to week 6 was 0.44 (95% confidence interval [CI], 0.15 to 0.73) in the 10-mg group (P=0.001), 0.49 (95% CI, 0.20 to 0.77) in the 50-mg group (P<0.001), and 0.48 (95% CI, 0.20 to 0.77) in the 100-mg group (P<0.001). The estimated differences from placebo in the change in intraepithelial lymphocyte density were -2.7 cells per 100 epithelial cells (95% CI, -7.6 to 2.2) in the 10-mg group, -4.2 cells per 100 epithelial cells (95% CI, -8.9 to 0.6) in the 50-mg group, and -9.6 cells per 100 epithelial cells (95% CI, -14.4 to -4.8) in the 100-mg group. Use of the 100-mg dose may have improved symptom and quality-of-life scores. The most common adverse events, the incidences of which were similar across all groups, were headache, nausea, diarrhea, vomiting, and abdominal pain. Rash developed in 3 of 40 patients (8%) in the 100-mg group. CONCLUSIONS In this preliminary trial, treatment with ZED1227 attenuated gluten-induced duodenal mucosal damage in patients with celiac disease.publishedVersionPeer reviewe

Serologic Assay for Diagnosis of Celiac Disease Based on a Patient-Derived Monoclonal Antigliadin Antibody

Manuscript title Serodiagnostic of celiac disease: Patient derived monoclonal anti-gliadin antibody harnessed in a novel inhibition assay Background & Aims Patients with celiac disease can be identified based on the detection of serum antibodies to deamidated gliadin peptides (DGPs). Recombinant human monoclonal antibodies (hmAb) against gliadin are produced by cloning antibody genes from single IgA-producing plasma cells isolated from lesions of patients with celiac disease. We developed an assay to identify patients with celiac disease based on the ability of antibodies from their serum to inhibit the binding of a gliadin-specific hmAb (1002-1E03) to a specific peptide antigen (inhibition assay). Methods We selected 2 peptides (a 34-mer and a 26-mer) found in ω-gliadins and low-molecular-weight glutenins that had been identified as specific targets of the hmAb 1002-1E03 from a digest of gliadin treated by transglutaminase 2. These peptides contained repeat sequence motifs; their interaction with hmAb 1002-1E03 was assessed in an amplified luminescent proximity homogeneous inhibition assay. We also tested peptides we created that included 3 repeated sequence motifs. Serum samples from untreated patients diagnosed with celiac disease (n = 106) and 2 control groups (198 blood donors, 151 patients with Crohn’s disease) were analyzed using the assay, as well as in conventional commercial assays that measure IgA against transglutaminase 2 (TG2) or IgG against DGP. Results In our inhibition assays, the 34-mer peptide showed the best results, and identified patients with celiac disease with 86.8% sensitivity and 98.6% specificity. Its diagnostic accuracy was comparable with that of commercial anti-DGP IgG (sensitivity, 87.9%; specificity, 98.0) and anti-TG2 IgA (sensitivity, 81.1%; specificity, 98.9) assays, and it detected most of the patients with anti-TG2 IgA-negative celiac disease without a significant decrease in specificity. Combined use of the anti-ω34 and the anti-TG2 assays produced specificity and sensitivity values of 95.3% and 98.0%, respectively. Conclusions We developed an antigliadin inhibition assay that identifies patients with celiac disease with high levels of specificity and sensitivity. It may prove useful as an adjunct to the current assay for anti-TG2 IgA