title: Human hepatocytes express trifluoroacetylated neoantigens after in vitro exposure to halothane.
abstract: Biotransformation of anaesthetic halothane by cytochrome P450-dependent monooxygenases resulted in the production of reactive intermediate trifluoroacetyl (TFA) halide, capable of covalently binding to hepatocyte proteins. TFA-modified liver proteins can act as antigens and are implicated in the pathogenesis of halothane hepatitis in humans. The aim of this study was to investigate the formation of TFA-neoantigens in halothane-treated primary cultures of adult human hepatocytes and to evaluate the usefulness of this in vitro model for studying immune-mediated halothane hepatotoxicity. Cultured human hepatocytes were incubated with halothane under constant temperature, atmosphere and anaesthetic concentration conditions. The results obtained show that halothane-treated hepatocytes isolated from seven different donors produced TFA-antigens as detected by immunocytochemical and western immunoblot analysis using rabbit anti-TFA antiserum. TFA-adducts were localized mainly in the endoplasmic reticulum and in small amounts on the plasma membrane of parenchymal cells. By immunoblotting, several neoantigens, with molecular masses from 42 to 100 kDa, were detected in halothane-exposed hepatocytes. These observations are consistent with the formation of TFA-adducts through metabolism of the anaesthetic and suggest that primary cultures of human hepatocytes represent a suitable in vitro model to study the pathogenesis of immune-mediated halothane hepatotoxicity.
authors.affiliation: INSERM U 49, Unité de Recherches Hépatologiques, Hôpital de Pontchaillou, Rennes, France.
authors.forename: G P
authors.initials: GP
authors.lastname: Ilyin
authors.forename: M
authors.initials: M
authors.lastname: Rissel
authors.forename: Y
authors.initials: Y
authors.lastname: Malledant
authors.forename: M
authors.initials: M
authors.lastname: Tanguy
authors.forename: A
authors.initials: A
authors.lastname: Guillouzo
mesh_terms: D000918:Antibody Specificity; D000941:Antigens / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology; D001711:Biotransformation; D002478:Cells, Cultured; D056486:Chemical and Drug Induced Liver Injury / Q000209:etiology / Q000276:immunology; D005463:Fluoroacetates; D006221:Halothane / Q000009:adverse effects / Q000494:pharmacology*; D006801:Humans; D008099:Liver / Q000187:drug effects* / Q000276:immunology / Q000648:ultrastructure; D012709:Serum Albumin / Q000276:immunology; D014269:Trifluoroacetic Acid / Q000276:immunology
chemical_list: D000941:Antigens; D005463:Fluoroacetates; D012709:Serum Albumin; C053344:trifluoroacetyl-serum albumin; D014269:Trifluoroacetic Acid; D006221:Halothane