title: An in vitro model of human acute ethanol exposure that incorporates CXCR3- and CXCR4-dependent recruitment of immune cells.
abstract: Alcoholic liver disease (ALD) is one of the commonest causes of cirrhosis and liver failure in the developed world. Hepatic inflammation is the critical stage in progression of both ALD and non-ALD, but it remains difficult to study the underlying mechanisms in a human system, and current animal models do not fully recapitulate human liver disease. We developed a human tissue-based system to study lymphocyte recruitment in response to ethanol challenge. Precision-cut liver slices (PCLS) from human livers were incubated in culture, and hepatic function was determined by albumin production, 3-(4,5-dimethylthiazol)-2,5-diphenyl tetrazolium bromide assay, glucose uptake responses, and morphometric assessment. Responses of tissue and lymphocytes to ethanol exposure were determined by PCR, flow cytometry, histology, and lymphocyte infiltration assays. Human PCLS demonstrated appropriate upregulation of CYP2E1, ADH1α, and ADH3 in response to ethanol treatment. Ethanol also induced expression of endothelial VCAM-1 and ICAM-1, production of sICAM-1 and CXCL8, and the chemokine receptors CXCR3 and CXCR4 on CD4 and CD8 lymphocytes. CXCR3- and CXCR4-dependent migration of lymphocytes into the tissue increased significantly in response to treatment with ethanol. We have demonstrated that ethanol increases chemokine receptor expression and lymphocyte recruitment into human liver tissue, suggesting that it may operate directly to promote hepatitis in ALD. The physiological and pathophysiological responses of the PCLS to ethanol in vitro highlight the potential of this assay for dissecting the molecular mechanisms underlying human liver inflammation and as a screening tool for novel therapeutics.
authors.affiliation: Institute of Biomedical Research, University of Birmingham, Birmingham, UK.
authors.forename: Sumera
authors.initials: S
authors.lastname: Karim
authors.forename: Evaggelia
authors.initials: E
authors.lastname: Liaskou
authors.forename: Samuel
authors.initials: S
authors.lastname: Hadley
authors.forename: Janine
authors.initials: J
authors.lastname: Youster
authors.forename: Jeff
authors.initials: J
authors.lastname: Faint
authors.forename: David H
authors.initials: DH
authors.lastname: Adams
authors.forename: Patricia F
authors.initials: PF
authors.lastname: Lalor
mesh_terms: D001483:Base Sequence; D002448:Cell Adhesion; D017931:DNA Primers; D000431:Ethanol / Q000633:toxicity*; D006801:Humans; D066298:In Vitro Techniques; D008214:Lymphocytes / Q000166:cytology / Q000187:drug effects*; D008954:Models, Biological; D016133:Polymerase Chain Reaction; D054367:Receptors, CXCR3 / Q000502:physiology*; D019718:Receptors, CXCR4 / Q000502:physiology*
chemical_list: C515187:CXCR3 protein, human; C527276:CXCR4 protein, human; D017931:DNA Primers; D054367:Receptors, CXCR3; D019718:Receptors, CXCR4; D000431:Ethanol