D. Ponticelli

Immunoregulatory Pathways Are Active in the Small Intestinal Mucosa of Patients with Potential Celiac Disease

OBJECTIVES:Potential celiac disease (CD) relates to subjects with a normal small intestinal mucosa who are at increased risk of developing CD as indicated by positive CD-associated serology. The objective of this study was to investigate in the small intestinal mucosa of such patients the state of immunological activation with special emphasis on immunoregulatory circuits.METHODS:Duodenal biopsies from active CD (n=48), potential CD (n=58), and control patients (n=45) were studied. RNA expression for interferon γ (IFNγ) and interleukin-10 (IL-10) were quantified by real-time quantitative PCR. The percentage of CD4+CD25+Foxp3+ T regulatory cells (Foxp3+Tregs) was determinated by flow cytometry and the number of Foxp3+ and IL-15+ cells by immunohistochemistry. Furthermore, we analyzed the suppressive function of CD4+CD25+ T cells, isolated from potential CD biopsy samples, as well as the effect of IL-15, on autologous peripheral blood responder CD4+CD25– T cells.RESULTS:In potential CD patients with Marsh 1 lesion, IFNγ-RNA expression was significantly less than in active, but enhanced if compared with potential CD patients with Marsh 0 lesion and with controls (P<0.001). The number of IL-15+ cells in subjects with potential CD was increased in comparison with controls (P<0.05), but lower than active CD (P<0.01). IL-10-RNA expression was upregulated in Marsh 0 potential CD patients if compared with those with Marsh 1 lesion (P<0.01) and controls (P<0.001), whereas there were no differences with active CD. The ratio IL-10/IFNγ reached the highest value in Marsh 0 potential CD compared with the other groups (P<0.05). The percentage of Foxp3+Tregs was also higher in potential CD compared with controls (P<0.05), although it was lower than in active CD (P<0.01). In co-culture assay, intestinal CD4+CD25+ T cells from potential CD patients exerted suppressive effects on T responder cells, and their activity was not impaired by IL-15.CONCLUSIONS:Potential CD patients show a low grade of inflammation that likely could be due to active regulatory mechanisms preventing the progression toward a mucosal damage.

In the Intestinal Mucosa of Children With Potential Celiac Disease IL-21 and IL-17A are Less Expressed than in the Active Disease

Objectives:Potential celiac disease (CD) patients are at an increased risk to developing CD as indicated by positive CD-associated serology. We investigated in duodenal mucosa of such patients the presence of both IL-21 and IL-17A and the role of gliadin peptides and IL-15 in their expression.Methods:Duodenal biopsies from 76 active CD, 90 potential CD, and 58 control patients were analyzed for IL-21 and/or IL-17A production by quantitative real-time PCR, immunohistochemistry, flow cytometry, and ELISA. The presence of IL-21 receptor was investigated by western blot. Potential CD duodenal fragments were cultured with gliadin peptides (PTG) and/or IL-15 and the expression/production of IL-21 and IL-17A assessed by quantitative real-time PCR and by immunohistochemistry.Results:In potential CD, IL-21 was lower than in active CD, in terms of RNA expression (P<0.01), density of lamina propria (LP) IL-21+ cells (P<0.05), and protein secretion (P<0.05). Also, IL-21R was weakly detectable in potential CD. Several LP cell types produced IL-21 in CD. In potential CD, CD4+IL-21+ cells increased after PMA-ionomycin stimulation and co-produced IFN-γ but not IL-17A. After 24 hours of culture stimulation with PTG, IL-21-producing cells increased but not the ones producing IL-17A. This increase was further enhanced by the addition of IL-15 to culture medium.Conclusions:In potential CD, IL-21 is less expressed than in active CD; however, IL-21-producing cells are present and prone to respond after specific stimuli. This suggests a key role of IL-21 in the progression of mucosal damage in CD.

Kinetics of anti-TG2 antibodies release into organ culture supernatants in celiac disease

that could not be clearly evaluated after GFD. Ultimately, 13/17 patients that received diagnosis without DB were aged <4 years (76% with median age 1.76). Among 114 patients that performed DB only 2 resulted in normal histology (potential CD) with a score of 1 and 3. Sixty out of 137 performed HLA: 51 of them had DQ2, 5 had DQ8 and 4 had both DQ2/DQ8. Associated disease were: 4 Hashimoto Tyroiditis, 1 Alopecia, 1 IDDM, 1 IgA deficit, 1 Pervasive developmental disorder. Conclusions:After two years of use, we can say that it should be very cautious in application of the new diagnostic criteria for CD. Great accuracy needs in evaluation of symptoms, especially those gluten-dependent, more convincing in subsequent confirmation of diagnosis. Sometimes are not enough, very high Ab levels alone. In our series the majority of patients had a score <4 and those with score like or higher than 4 are usually very young. This approach canbe aid to assist in the evaluationof theweight of eachdiagnostic sign and toprotect against overdiagnosis innotwell documentedor borderline cases. Moreover, potential CD patients are rare (2/114) but this, probably results by our habit not to submit soon to DB children with Ab titer <3 times of cut-off value.