Anderson and co-workers established an innovative approach that allows the detection BAY 80-6946 in vitro of gluten-specific T cells in the peripheral blood of CD patients after a short period of gluten-containing food consumption [4,5]. Basically, gluten-sensitized

CD4+ T cells, normally scarcely detectable in the blood of coeliac patients, circulate transiently in the peripheral blood after 3 days of wheat challenge, and can be detected by a sensitive interferon (IFN)-γ enzyme-linked inmmunospot (ELISPOT) assay. Using this in-vivo procedure, the authors further screened large libraries of prolamin peptides and assessed the hierarchy and immunodominance of gluten T cell epitopes [6]. More recently, T cells reactive to DQ2-α-I and DQ2-α-II epitopes were monitored in the peripheral blood of bread-challenged coeliacs with specific DQ2-tetramer constructs [7,8]. Of BAY 73-4506 clinical trial note, both Australian and Norwegian studies enrolled adult coeliac volunteers, with an average age of 43 years. To our knowledge, no information is available on the responsiveness to short gluten challenge in very young coeliac patients. Furthermore, very little is known about the in-vivo challenge reproducibility

in the same subject cohort, with the exception of a few cases of coeliacs who underwent two separate gluten consumptions described in the above-mentioned studies [7,8]. In the present study we have validated the in-vivo short gluten challenge in a cohort of

14 young CD patients of Italian origin. In particular, we analysed the peripheral blood response against whole gliadin and the immunodominant 33-mer peptide (α-gliadin 57–89). We also assessed the feasibility of exposing the patient cohort to a second in-vivo challenge after a period of 3–10 months of wash-out, in order to estimate the reproducibility of the procedure in the same study population and the intra-individual variations. If replicated successfully in other studies, the short wheat challenge could Fluorouracil concentration represent a strategic tool to evaluate non-invasively the individual’s response to gluten, and could be applied to intervention studies. In fact, the evaluation of small bowel mucosa damage after long-term wheat challenge has been used since the 1950s to assess cereal toxicity or to define the toxic peptides [9–11]. Such studies required repeated endoscopies, before and after treatment, which are always not well accepted by participants. To detect a dysregulated response to gluten, other functional markers, such as faecal fat and xylose malabsorption, resulted in low specificity and sensitivity [12–15]. Furthermore, recent studies have indicated that a short gluten challenge could be used to support diagnosis in doubtful cases of CD [16–18]. Fourteen DQ2-positive volunteers with CD (mean age 18·6, range 15–24 years) participated in the study (Table 1).