title: Development of a modified lymphocyte transformation test for diagnosing drug-induced liver injury associated with an adaptive immune response.
abstract: Drug-induced liver injury (DILI) is a growing problem. Diagnostic methods to differentiate DILI caused by an adaptive immune response from liver injury of other causes or to identify the responsible drug in patients receiving multiple drugs, herbals and/or dietary supplements (polypharmacy) have not yet been established. The lymphocyte transformation test (LTT) has been proposed as a diagnostic method to determine if a subject with an apparent hypersensitivity reaction has become sensitized to a specific drug. In this test, peripheral blood mononuclear cells (PBMC) collected from a subject are incubated with drug(s) suspected of causing the reaction. Cell proliferation, measured by the incorporation of [3H]-thymidine into new DNA, is considered evidence of a drug-specific immune response. The objectives of the current studies were to: (1) develop and optimize a modified version of the LTT (mLTT) and (2) investigate the feasibility of using the mLTT for diagnosing DILI associated with an adaptive immune response and identifying the responsible drug. PBMC collected from donors with a history of drug hypersensitivity reactions to specific drugs (manifested as skin rash) were used as positive controls for assay optimization. Following optimization, samples collected from 24 subjects enrolled in the U.S. Drug-Induced Liver Injury Network (DILIN) were tested in the mLTT. Using cytokine and granzyme B production as the primary endpoints to demonstrate lymphocyte sensitization to a specific drug, most samples from the DILIN subjects failed to respond. However, robust positive mLTT responses were observed for two of four samples from three DILIN subjects with hepatitis due to isoniazid (INH). We conclude that the mLTT, as performed here on frozen and thawed PBMC, is not a reliable test for diagnosing DILI caused by all drugs, but that it may be useful for confirming the role of the adaptive immune response in DILI ascribed to INH.
authors.affiliation: a Drug Safety Research and Development, Pfizer Inc. , Groton , CT , USA.
authors.forename: Jessica
authors.initials: J
authors.lastname: Whritenour
authors.affiliation: b Drug Safety Research and Development, Pfizer Inc. , La Jolla , CA , USA.
authors.forename: Mira
authors.initials: M
authors.lastname: Ko
authors.affiliation: b Drug Safety Research and Development, Pfizer Inc. , La Jolla , CA , USA.
authors.forename: Qing
authors.initials: Q
authors.lastname: Zong
authors.affiliation: b Drug Safety Research and Development, Pfizer Inc. , La Jolla , CA , USA.
authors.forename: Jianying
authors.initials: J
authors.lastname: Wang
authors.affiliation: a Drug Safety Research and Development, Pfizer Inc. , Groton , CT , USA.
authors.forename: Karrie
authors.initials: K
authors.lastname: Tartaro
authors.affiliation: a Drug Safety Research and Development, Pfizer Inc. , Groton , CT , USA.
authors.forename: Patricia
authors.initials: P
authors.lastname: Schneider
authors.affiliation: a Drug Safety Research and Development, Pfizer Inc. , Groton , CT , USA.
authors.forename: Ellen
authors.initials: E
authors.lastname: Olson
authors.affiliation: a Drug Safety Research and Development, Pfizer Inc. , Groton , CT , USA.
authors.forename: Maria
authors.initials: M
authors.lastname: Van Volkenburg
authors.affiliation: c National Institute of Diabetes and Digestive and Kidney Diseases, NIH , Bethesda , MD , USA.
authors.forename: Jose
authors.initials: J
authors.lastname: Serrano
authors.affiliation: d Department of Internal Medicine , University of North Carolina , Chapel Hill , NC , USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Hayashi
authors.affiliation: e Department of Internal Medicine , University of Michigan , Ann Arbor , MI , USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Fontana
authors.affiliation: f Department of Internal Medicine , IUPUI , Indianapolis , IN , USA.
authors.forename: Naga
authors.initials: N
authors.lastname: Chalasani
authors.affiliation: g Department of Internal Medicine , Wake Forest University School of Medicine , Winston-Salem , NC , USA.
authors.forename: Herbert L
authors.initials: HL
authors.lastname: Bonkovsky
mesh_terms: D000208:Acute Disease; D056704:Adaptive Immunity; D049109:Cell Proliferation; D002478:Cells, Cultured; D056486:Chemical and Drug Induced Liver Injury / Q000175:diagnosis* / Q000276:immunology; D016207:Cytokines / Q000378:metabolism; D003937:Diagnosis, Differential; D004342:Drug Hypersensitivity / Q000175:diagnosis* / Q000276:immunology; D005240:Feasibility Studies; D005500:Follow-Up Studies; D053804:Granzymes / Q000378:metabolism; D006505:Hepatitis / Q000175:diagnosis* / Q000276:immunology; D006801:Humans; D007159:Immunologic Tests / Q000379:methods*; D007538:Isoniazid / Q000009:adverse effects* / Q000627:therapeutic use; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000276:immunology*; D008213:Lymphocyte Activation
chemical_list: D016207:Cytokines; D053804:Granzymes; D007538:Isoniazid
keywords: Allergic reactions; drug-induced liver injury; drugs; hepatitis; immuno-allergic; lymphocyte transformation test; lymphocytes