title: Detection of Primary T Cell Responses to Drugs and Chemicals in HLA-Typed Volunteers: Implications for the Prediction of Drug Immunogenicity.
abstract: A number of serious adverse drug reactions are caused by T cells. An association with HLA alleles has been identified with certain reactions, which makes it difficult to develop standardized preclinical tests to predict chemical liability. We have recently developed a T cell priming assay using the drug metabolite nitroso sulfamethoxazole (SMX-NO). We now report on reproducibility of the assay, establishment of a biobank of PBMC from 1000 HLA-typed volunteers, and generation of antigen-specific responses to a panel of compounds. Forty T cell priming assays were performed with SMX-NO; 5 gave weak responses (1.5-1.9) and 34 showed good (SI 2.0-3.9) or strong responses (SI  > 4.0) using readouts for proliferation and cytokine release. Thus, SMX-NO can be used as a model reagent for in vitro T cell activation. Good to strong responses were also generated to haptenic compounds (amoxicillin, piperacillin and Bandrowski's base) that are not associated with an HLA risk allele. Furthermore, responses were detected to carbamazepine (in HLA-B*15:02 donors), flucloxacillin (in 1 HLA-B*57:01 donor) and oxypurinol (in HLA-B*58:01 donors), which are associated with HLA-class I-restricted forms of hypersensitivity. In contrast, naïve T cell priming to ximelagatran, lumiracoxib, and lapatinib (HLA-class II-restricted forms of hypersensitivity) yielded negative results. Abacavir, which activates memory T cells in patients, did not activate naïve T cells from HLA-B*57:01 donors. This work shows that the priming assay can be used to assess primary T cell responses to drugs and to study mechanisms T cell priming for drugs that display HLA class I restriction. Additional studies are required to investigate HLA-class II-restricted reactions.
authors.affiliation: Department of Molecular and Clinical Pharmacology, Centre for Drug Safety Science; bkpark@liverpool.ac.uk.
authors.forename: Lee
authors.initials: L
authors.lastname: Faulkner
authors.affiliation: Department of Molecular and Clinical Pharmacology, Centre for Drug Safety Science; bkpark@liverpool.ac.uk.
authors.forename: Andrew
authors.initials: A
authors.lastname: Gibson
authors.affiliation: Department of Molecular and Clinical Pharmacology, Centre for Drug Safety Science; bkpark@liverpool.ac.uk.
authors.forename: Andrew
authors.initials: A
authors.lastname: Sullivan
authors.affiliation: Department of Molecular and Clinical Pharmacology, Centre for Drug Safety Science; bkpark@liverpool.ac.uk.
authors.forename: Arun
authors.initials: A
authors.lastname: Tailor
authors.affiliation: Department of Molecular and Clinical Pharmacology, Centre for Drug Safety Science; bkpark@liverpool.ac.uk.
authors.forename: Toru
authors.initials: T
authors.lastname: Usui
authors.affiliation: Department of Personalised Medicine, Institute of Translational Medicine, University of Liverpool, Liverpool, UK bkpark@liverpool.ac.uk.
authors.forename: Ana
authors.initials: A
authors.lastname: Alfirevic
authors.affiliation: Department of Personalised Medicine, Institute of Translational Medicine, University of Liverpool, Liverpool, UK bkpark@liverpool.ac.uk.
authors.forename: Munir
authors.initials: M
authors.lastname: Pirmohamed
authors.affiliation: Department of Molecular and Clinical Pharmacology, Centre for Drug Safety Science; bkpark@liverpool.ac.uk.
authors.forename: Dean J
authors.initials: DJ
authors.lastname: Naisbitt
authors.affiliation: Department of Molecular and Clinical Pharmacology, Centre for Drug Safety Science; bkpark@liverpool.ac.uk.
authors.forename: B
authors.initials: B
authors.lastname: Kevin Park
mesh_terms: D000328:Adult; D018070:Biological Specimen Banks; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D018920:Coculture Techniques; D016207:Cytokines / Q000378:metabolism; D004342:Drug Hypersensitivity / Q000276:immunology* / Q000378:metabolism; D005260:Female; D005787:Gene Frequency; D006680:HLA Antigens / Q000235:genetics / Q000276:immunology*; D006650:Histocompatibility Testing; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D008213:Lymphocyte Activation / Q000187:drug effects*; D008297:Male; D018570:Risk Assessment; D013420:Sulfamethoxazole / Q000031:analogs & derivatives* / Q000633:toxicity; D013601:T-Lymphocytes / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D018675:Toxicity Tests / Q000379:methods*; D055815:Young Adult
chemical_list: D016207:Cytokines; D006680:HLA Antigens; C072956:4-nitrososulfamethoxazole; D013420:Sulfamethoxazole
keywords: HLA; T cells; drug hypersensitivity.