title: Patients with inflammatory bowel disease (IBD) reveal increased induction capacity of intracellular interferon-gamma (IFN-gamma) in peripheral CD8+ lymphocytes co-cultured with intestinal epithelial cells.
abstract: Intestinal epithelial cells seem to play a key role during IBD. The network of cellular interactions between epithelial cells and lamina propria mononuclear cells is still incompletely understood. In the following co-culture model we investigated the influence of intestinal epithelial cells on cytokine expression of T cytotoxic and T helper cells from patients with IBD and healthy controls. Peripheral blood mononuclear cells (PBMC) were purified by a Ficoll-Hypaque gradient followed by co-incubation with epithelial cells in multiwell cell culture insert plates in direct contact as well as separated by transwell filters. We used Caco-2 cells as well as freshly isolated colonic epithelia obtained from surgical specimens. Three-colour immunofluorescence flow cytometry was performed after collection, stimulation and staining of PBMC with anti-CD4, anti-CD8, anti-IFN-gamma and anti-IL-4. Patients with IBD (Crohn's disease (CD), n = 12; ulcerative colitis (UC), n = 16) and healthy controls (n = 10) were included in the study. After 24 h of co-incubation with Caco-2 cells we found a significant increase of IFN-gamma-producing CD8+ lymphocytes in patients with IBD. In contrast, healthy controls did not respond to the epithelial stimulus. No significant differences could be found between CD and UC or active and inactive disease. A significant increase of IFN-gamma+/CD8+ lymphocytes in patients with UC was also seen after direct co-incubation with primary cultures of colonic crypt cells. The observed epithelial-lymphocyte interaction seems to be MHC I-restricted. No significant epithelial cell-mediated effects on cytokine expression were detected in the PBMC CD4+ subsets. Patients with IBD-even in an inactive state of disease-exert an increased capacity for IFN-gamma induction in CD8+ lymphocytes mediated by intestinal epithelial cells. This mechanism may be important during chronic intestinal inflammation, as in the case of altered mucosal barrier function epithelial cells may become targets for IFN-gamma-producing CD8+ lymphocytes.
authors.affiliation: Department of Medicine B, Department of General Surgery and Institute of Immunology, University of Münster, Münster, Germany.
authors.forename: G
authors.initials: G
authors.lastname: Bisping
authors.forename: N
authors.initials: N
authors.lastname: Lügering
authors.forename: S
authors.initials: S
authors.lastname: Lütke-Brintrup
authors.forename: H G
authors.initials: HG
authors.lastname: Pauels
authors.forename: G
authors.initials: G
authors.lastname: Schürmann
authors.forename: W
authors.initials: W
authors.lastname: Domschke
authors.forename: T
authors.initials: T
authors.lastname: Kucharzik
mesh_terms: D019138:Antibodies, Blocking / Q000494:pharmacology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology* / Q000378:metabolism*; D018938:Caco-2 Cells; D002478:Cells, Cultured; D018920:Coculture Techniques; D003093:Colitis, Ulcerative / Q000276:immunology / Q000473:pathology; D003106:Colon; D005260:Female; D015395:Histocompatibility Antigens Class I / Q000276:immunology; D006801:Humans; D007106:Immune Sera / Q000494:pharmacology; D015212:Inflammatory Bowel Diseases / Q000276:immunology* / Q000473:pathology; D007371:Interferon-gamma / Q000096:biosynthesis* / Q000097:blood; D015847:Interleukin-4 / Q000096:biosynthesis; D007413:Intestinal Mucosa / Q000166:cytology / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D007424:Intracellular Fluid / Q000276:immunology* / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000276:immunology / Q000378:metabolism; D008297:Male; D009000:Monocytes / Q000276:immunology; D015398:Signal Transduction / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology / Q000378:metabolism
chemical_list: D019138:Antibodies, Blocking; D015395:Histocompatibility Antigens Class I; D007106:Immune Sera; D015847:Interleukin-4; D007371:Interferon-gamma