title: Standardization of a human organ culture model of intestinal inflammation and its application for drug testing.
abstract: Targeting early molecular events in intestinal inflammation may represent a useful therapeutic strategy for maintaining remission in inflammatory bowel disease. Recently, we established an intestinal organ culture model (LEL model), which allows to study the initiation of an intestinal inflammatory response in human tissue. In this model, EDTA-mediated depletion of epithelial cells of colonic mucosa results in an instantaneous inflammatory response in resident lamina propria cells, which shows features of intestinal inflammation in vivo. Furthermore, activated immune cells emigrate from the lamina propria onto the luminal side of the basement membrane. Here, we standardize the LEL model and explore its suitability for drug testing. To this end, human mucosal punches of defined surface area were prepared, depleted of epithelial cells, and cultured at an optimized ratio of medium volume/punch area. The intra-assay variability of measurements of inflammatory parameters ranged from 13% for cell migration to 19% for secretion and 30% for tissue gene expression, respectively, of the inflammatory mediators IL-8 and IL-6. Importantly, known suppressive effects of dexamethasone, a drug employed for the treatment of inflammatory bowel diseases, on leucocyte migration, IL8, IL6, and TNF-α production as well as CD86 surface expression by myeloid cells were observed in this model. In conclusion, the present results suggest that the LEL model may represent a useful human experimental system not only for studying initial activation mechanisms in intestinal inflammation but also for evaluating drug compounds for the treatment of mucosal inflammation.
authors.affiliation: Institute of Immunology, University Hospital Heidelberg, INF 305, D-69120 Heidelberg, Germany; Department of Anesthesiology, University Hospital Heidelberg, INF 110, D-69120 Heidelberg, Germany.
authors.forename: Timea
authors.initials: T
authors.lastname: Szikszai
authors.affiliation: Institute of Pathology, University Hospital Heidelberg, INF 224, D-69120 Heidelberg, Germany.
authors.forename: Felix
authors.initials: F
authors.lastname: Lasitschka
authors.affiliation: Institute of Immunology, University Hospital Heidelberg, INF 305, D-69120 Heidelberg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Giese
authors.affiliation: Institute of Immunology, University Hospital Heidelberg, INF 305, D-69120 Heidelberg, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Greulich
authors.affiliation: Institute of Immunology, University Hospital Heidelberg, INF 305, D-69120 Heidelberg, Germany.
authors.forename: Young-Seon
authors.initials: YS
authors.lastname: Lee
authors.affiliation: Institute of Immunology, University Hospital Heidelberg, INF 305, D-69120 Heidelberg, Germany.
authors.forename: Juliane
authors.initials: J
authors.lastname: Ilse
authors.affiliation: Institute of Immunology, University Hospital Heidelberg, INF 305, D-69120 Heidelberg, Germany; Department of Surgery, University Hospital Heidelberg, INF 110, D-69120 Heidelberg, Germany.
authors.forename: Serin
authors.initials: S
authors.lastname: Schiessling
authors.affiliation: Department of Surgery, University Hospital Heidelberg, INF 110, D-69120 Heidelberg, Germany.
authors.forename: Christine
authors.initials: C
authors.lastname: Leowardi
authors.affiliation: Department of Surgery, University Hospital Heidelberg, INF 110, D-69120 Heidelberg, Germany.
authors.forename: Mohammed
authors.initials: M
authors.lastname: Al-Saeedi
authors.affiliation: Department of Surgery, St. Vincentius Hospital, Holzstr. 4a, D-67346 Speyer, Germany.
authors.forename: Johannes
authors.initials: J
authors.lastname: Winter
authors.affiliation: Institute of Immunology, University Hospital Heidelberg, INF 305, D-69120 Heidelberg, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Meuer
authors.affiliation: Institute of Immunology, University Hospital Heidelberg, INF 305, D-69120 Heidelberg, Germany. Electronic address: jutta.schroeder-braunstein@immu.uni-heidelberg.de.
authors.forename: Jutta
authors.initials: J
authors.lastname: Schröder-Braunstein
mesh_terms: D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D051940:B7-2 Antigen / Q000096:biosynthesis; D002465:Cell Movement / Q000276:immunology; D003106:Colon / Q000166:cytology / Q000276:immunology* / Q000473:pathology; D003907:Dexamethasone / Q000494:pharmacology*; D006801:Humans; D007249:Inflammation / Q000276:immunology; D015212:Inflammatory Bowel Diseases / Q000276:immunology* / Q000473:pathology; D015850:Interleukin-6 / Q000096:biosynthesis / Q000378:metabolism; D016209:Interleukin-8 / Q000096:biosynthesis / Q000378:metabolism; D007413:Intestinal Mucosa / Q000166:cytology / Q000276:immunology / Q000473:pathology; D022423:Myeloid Cells / Q000276:immunology; D009924:Organ Culture Techniques / Q000379:methods*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000378:metabolism
chemical_list: D000893:Anti-Inflammatory Agents; D051940:B7-2 Antigen; C497964:CD86 protein, human; C508606:CXCL8 protein, human; C508600:IL6 protein, human; D015850:Interleukin-6; D016209:Interleukin-8; D014409:Tumor Necrosis Factor-alpha; D003907:Dexamethasone
keywords: Drug testing; Human organ culture; Intestinal inflammation; Standardization