In vitro Model of the Pathogenesis of Celiac Disease

Abstract

The in vitro challenge of duodenal mucosa with gliadin is a useful model to reproduce the immunological features of celiac disease (CD) and allows the study of early pathogenetic events in this disease. With this model it was shown that antigens such as ICAM-1 and HLA-DR are upregulated as early as 1–2 h after gliadin challenge in patients with CD. After 24 h the lamina propria contained CD4+ T cells expressing the IL-2 receptor α-chain, which is a sign of activation. Intraepithelial lymphocytes increased in number and showed proliferative activity. After in vitro stimulation with gliadin, endomysial antibodies were found in the supernatant of the cultured mucosa from patients with CD following a gluten-free diet. This supported the notion that endomysial antibodies are at least in part produced locally. The model was also successfully used to identify toxic constituents of gliadin. Presently, organ culture is not commonly used for diagnostic purposes.

In Vitro Gluten Challenge Test for Celiac Disease Diagnosis

“…Ã One study evaluated patients with partial GFD (37). y One study evaluated specimens at 72 hours after challenge (14). dependent.…”

“…In the in vitro method, antigen markers such as intercellular cell adhesion molecule (ICAM)-1 and HLA-DR, CD markers, histological findings, EMA-immunoglobulin A (IgA), and anti-tTG levels are evaluated in the supernatant of the 3 mucosa culture from patients suspected of having CD after a gluten challenge test (14). IgA EMAs were searched in undiluted culture supernatants by indirect immunofluorescence analysis on the cryostat sections of primate esophagus.…”

Identification of a new coeliac disease subgroup: antiendomysial and anti‐transglutaminase antibodies of IgG class in the absence of selective IgA deficiency

“…In fact, in vitro studies demonstrated that intestinal mucosa of treated CD patients in clinical remission is sensitive to gluten. It has been demonstrated that biopsy culture in the presence of gliadin is able to reproduce features of immunological activation typical of CD [4, 20]. In addition, a gluten‐dependent tissue immunological activation in the absence of histological damage has already been described [5, 21].…”

Increased Stability of Glycol-Terminated Self-Assembled Monolayers for Long-Term Patterned Cell Culture

“…Classical systems for patterned cell culture, including self-assembled monolayers (SAMs) formed from alkanethiols on gold, have limited stability under cell culture conditions. Most systems are only stable for 5–7 days in cell culture, , which significantly limits their use for the study of developmental events, in vitro disease models, and long-term model systems for drug discovery. − Monolayer instability has limited the use of patterned substrates to short-term cell culture experiments lasting only 1–2 days. ,,, Here, we develop a system that is stable for over 5 weeks in culture and explore the mechanism of SAM degradation, which has been of some debate.…”