Celiac disease (CD) is an inflammatory bowel disease triggered by gluten in genetically susceptible individuals (HLA-DQ2/DQ8). It leads to duodenal histological alterations and specific serum autoantibody response, with different clinical manifestations. CD has a high prevalence (1%) and may involve any age.
Extracellular microvesicles (MVs) contain a broad spectrum of bioactive substances (signaling proteins, mRNA, micro RNA, bioactive lipids) and are released in body fluids by different cell types under stress, injury and inflammatory conditions. MVs produced by intestinal epithelial cells are implicated in maintaining homeostasis and in the spread of inflammation. In physiological conditions, intestinal epithelial cells release MVs from their apical surface in the lumen. Furthermore, MVs released into the mucosa may carry antigens to dendritic cells regulating the adaptative immune response. In pathological condition, MVs isolated from active IBD patients showed significantly higher content of inflammatory cytokines, mRNA and protein levels than healthy controls. Furthermore, the uptake of such MVs by epithelial cells led to increased pro-inflammatory cytokine expression and induction of macrophage migration, suggesting that MVs may actively contribute to local propagation of inflammation.
The aim of this study is to evaluate the role of MVs, purified from plasma and duodenal biopsy cultures of CD patients, as possible biomarkers and pathogenic players in gut inflammation.
We will enroll 20 active CD patients, 20 CD patients in remission from at least 12 months and 20 control patients. Bioptic specimen supernatant and plasma MV purification will be performed, as well as proteomic analysis of purified MVs. Effects of MVs on Caco-2 cells will be studied by means of transepithelial electrical resistance, confocal laser scanning microscopy, cells apoptosis detection and Caco-2 cells peripheral blood co-culture.
Celiac disease (CD) has an estimated prevalence of 1%, its pathogenesis is welll known and specific and/or sensitive diagnostic methods are widely used. However, it is very important an early diagnosis to reduce both the large spectrum of comorbidity and costs associated with a late diagnosis.
Nowadays, a specific biomarker of early diagnosis and patient follow-up is still unknown. Current literature suggests that extracellular MVs may be a novel source of biomarkers in inflammatory diseases. On these basis, our aim will be to investigate the role of MVs purified from plasma and duodenal biopsy culture supernatant of CD patients (taken as gut inflammation model) using Caco-2 cell system, a well characterized intestinal in vitro model. The identification of new potential markers could increase knowledge of the pathogenic mechanisms of CD and could allow to save time and resources in CD case finding, especially when diagnosis is difficult to reach (e.g. seronegative patients).
All these laboratory results will be finally compared with a sophisticated nutritional assessment of all patients enrolled, to search for possible correlations with CD inflammatory state and purified MVs effects.