Article

Gliadin cytotoxicity and in vitro cell cultures.

Department of Biology and Genetics, University of Milan, Via Votti 9, Milan 20100, Italy.
Toxicology Letters (impact factor: 3.23). 01/2004; 146(1):1-8.
Source: PubMed

ABSTRACT Interest in celiac disease, a common enteropathy in Europe and the USA (1/200) caused by the dietary ingestion of gluten in susceptible subjects, has increased over the last few years. Its pathogenesis is still not completely clear, but it certainly involves immune-mediated mechanisms. Although a number of studies have been published concerning the role of T cells in inducing intestinal damage, little is known about the early stages in which gliadin (the toxic component of gluten) starts the whole process. In vitro two- and three-dimensional (multicellular spheroid) cell cultures are a simple and useful means of studying the direct cellular effects of gliadin and other "toxic" cereal peptides. Furthermore, in addition to improving our understanding of pathogenetic mechanisms, cell cultures can also be used to test modified peptides that could replace the toxic components present in the foods that celiac patients must avoid.

0 0
 · 
0 Bookmarks
 · 
24 Views
  • Source
    Article: Diabetes-specific HLA-DR-restricted proinflammatory T-cell response to wheat polypeptides in tissue transglutaminase antibody-negative patients with type 1 diabetes.
    Majid Mojibian, Habiba Chakir, David E Lefebvre, Jennifer A Crookshank, Brigitte Sonier, Erin Keely, Fraser W Scott
    [show abstract] [hide abstract]
    ABSTRACT: There is evidence of gut barrier and immune system dysfunction in some patients with type 1 diabetes, possibly linked with exposure to dietary wheat polypeptides (WP). However, questions arise regarding the frequency of abnormal immune responses to wheat and their nature, and it remains unclear whether such responses are diabetes specific. In type 1 diabetic patients and healthy control subjects, the immune response of peripheral CD3(+) T-cells to WPs, ovalbumin, gliadin, alpha-gliadin 33-mer peptide, tetanus toxoid, and phytohemagglutinin was measured using a carboxyfluorescein diacetate succinimidyl ester (CFSE) proliferation assay. T-helper cell type 1 (Th1), Th2, and Th17 cytokines were analyzed in WP-stimulated peripheral blood mononuclear cell (PBMNC) supernatants, and HLA was analyzed by PCR. Of 42 patients, 20 displayed increased CD3(+) T-cell proliferation to WPs and were classified as responders; proliferative responses to other dietary antigens were less pronounced. WP-stimulated PBMNCs from patients showed a mixed proinflammatory cytokine response with large amounts of IFN-gamma, IL-17A, and increased TNF. HLA-DQ2, the major celiac disease risk gene, was not significantly different. Nearly all responders carried the diabetes risk gene HLA-DR4. Anti-DR antibodies blocked the WP response and inhibited secretion of Th1 and Th17 cytokines. High amounts of WP-stimulated IL-6 were not blocked. T-cell reactivity to WPs was frequently present in type 1 diabetic patients and associated with HLA-DR4 but not HLA-DQ2. The presence of an HLA-DR-restricted Th1 and Th17 response to WPs in a subset of patients indicates a diabetes-related inflammatory state in the gut immune tissues associated with defective oral tolerance and possibly gut barrier dysfunction.
    Diabetes 05/2009; 58(8):1789-96. · 8.29 Impact Factor
  • Source
    Article: Isolation and culture of fibroblasts from endoscopic duodenal biopsies of celiac patients.
    Leda Roncoroni, Luca Elli, Luisa Doneda, Luca Piodi, Michele M Ciulla, Roberta Paliotti, Maria Teresa Bardella
    [show abstract] [hide abstract]
    ABSTRACT: Fibroblasts are actually considered pivotal in inflammation and tissue remodelling process and for these reasons they are involved in the pathogenesis of autoimmune disorders such as celiac disease. Investigations to define the role of fibroblasts in celiac diseases are obstructed by the absence of specific models. Our objective is to isolate and culture primary fibroblasts from endoscopic duodenal biopsies of celiac and non-celiac subjects, to analyze their growth patterns and the morphometric characteristics. 60 duodenal bioptic specimens from 20 celiac patients and 114 from 38 non-celiac subjects were mechanically chopped and enzymatically digested in order to obtain primary cell cultures. Growth patterns, karyotype (Q-banding analysis), expression of typing proteins (fibroblast surface protein and cytokeratin 20) and morphometric parameters (diameters and their ratio, perimeter, area and perimeter/area ratio at computerised image analysis) were investigated on cultured cells. Primary cells were successfully cultured in 78% of the collected duodenal biopsies. Cultured cells, expressing the fibroblast surface protein, were negative for cytokeratine 20 and maintained a normal kariotype. Cells grew slowly without differences between the celiac and the non celiac group. Morphometric analysis of celiac fibroblasts revealed significantly increased dimensions, with a preserved diameters ratio, and a reduced perimeter/area ratio. For the first time this study demonstrates the feasibility of culturing primary fibroblast cell from endoscopic duodenal biopsies in celiac and non-celiac subjects, opening a new window of opportunity in studies intended to establish the role of fibroblasts as a possible partaker in the pathogenesis of the celiac mucosal damage.
    Journal of Translational Medicine 02/2009; 7:40. · 3.41 Impact Factor
Data provided are for informational purposes only. Although carefully collected, accuracy cannot be guaranteed. The impact factor represents a rough estimation of the journal's impact factor and does not reflect the actual current impact factor. Publisher conditions are provided by RoMEO. Differing provisions from the publisher's actual policy or licence agreement may be applicable.

Keywords

celiac disease
 
celiac patients
 
cell cultures
 
cereal peptides
 
common enteropathy
 
dietary ingestion
 
direct cellular effects
 
gliadin
 
immune-mediated mechanisms
 
inducing intestinal damage
 
last
 
multicellular spheroid
 
pathogenetic mechanisms
 
susceptible subjects
 
T cells
 
three-dimensional
 
toxic component
 
toxic components present
 
vitro two-
 
whole process