title: Oxypurinol-Specific T Cells Possess Preferential TCR Clonotypes and Express Granulysin in Allopurinol-Induced Severe Cutaneous Adverse Reactions.
abstract: Allopurinol, a first-line drug for treating gout and hyperuricemia, is one of the leading causes of severe cutaneous adverse reactions (SCARs). To investigate the molecular mechanism of allopurinol-induced SCAR, we enrolled 21 patients (13 Stevens-Johnson syndrome (SJS)/toxic epidermal necrolysis (TEN) and 8 drug reaction with eosinophilia and systemic symptoms (DRESS)), 11 tolerant controls, and 23 healthy donors. We performed in vitro T-cell activation assays by culturing peripheral blood mononuclear cells (PBMCs) with allopurinol, oxypurinol, or febuxostat and measuring the expression of granulysin and IFN-γ in the supernatants of cultures. TCR repertoire was investigated by next-generation sequencing. Oxypurinol stimulation resulted in a significant increase in granulysin in the cultures of blood samples from SCAR patients (n=14) but not tolerant controls (n=11) or healthy donors (n=23). Oxypurinol induced T-cell response in a concentration- and time-dependent manner, whereas allopurinol or febuxostat did not. T cells from patients with allopurinol-SCAR showed no crossreactivity with febuxostat. Preferential TCR-V-β usage and clonal expansion of specific CDR3 (third complementarity-determining region) were found in the blister cells from skin lesions (n=8) and oxypurinol-activated T-cell cultures (n=4) from patients with allopurinol-SCAR. These data suggest that, in addition to HLA-B*58:01, clonotype-specific T cells expressing granulysin upon oxypurinol induction participate in the pathogenesis of allopurinol-induced SCAR.
authors.affiliation: Department of Dermatology, Drug Hypersensitivity Clinical and Research Center, Chang Gung Memorial Hospital, Linkou, Taipei and Keelung, Taiwan; College of Medicine, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Wen-Hung
authors.initials: WH
authors.lastname: Chung
authors.affiliation: Program in Molecular Medicine, National Yang-Ming University and Academia Sinica, Taipei, Taiwan.
authors.forename: Ren-You
authors.initials: RY
authors.lastname: Pan
authors.affiliation: Department and Institute of Pharmacology, National Yang-Ming University, Taipei, Taiwan.
authors.forename: Mu-Tzu
authors.initials: MT
authors.lastname: Chu
authors.affiliation: Department of Dermatology, Drug Hypersensitivity Clinical and Research Center, Chang Gung Memorial Hospital, Linkou, Taipei and Keelung, Taiwan; College of Medicine, Chang Gung University, Taoyuan, Taiwan.
authors.forename: See-Wen
authors.initials: SW
authors.lastname: Chin
authors.affiliation: Department of Dermatology, Drug Hypersensitivity Clinical and Research Center, Chang Gung Memorial Hospital, Linkou, Taipei and Keelung, Taiwan; College of Medicine, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Yu-Lin
authors.initials: YL
authors.lastname: Huang
authors.affiliation: Health GeneTech Corporation, Taoyuan, Taiwan.
authors.forename: Wei-Chi
authors.initials: WC
authors.lastname: Wang
authors.affiliation: Health GeneTech Corporation, Taoyuan, Taiwan.
authors.forename: Jen-Yun
authors.initials: JY
authors.lastname: Chang
authors.affiliation: Program in Molecular Medicine, National Yang-Ming University and Academia Sinica, Taipei, Taiwan; Department and Institute of Pharmacology, National Yang-Ming University, Taipei, Taiwan. Electronic address: sihung@ym.edu.tw.
authors.forename: Shuen-Iu
authors.initials: SI
authors.lastname: Hung
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000493:Allopurinol / Q000009:adverse effects* / Q000494:pharmacology; D000945:Antigens, Differentiation, T-Lymphocyte / Q000378:metabolism*; D016022:Case-Control Studies; D002478:Cells, Cultured / Q000276:immunology; D003429:Cross Reactions; D003875:Drug Eruptions / Q000209:etiology / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D000069465:Febuxostat; D005260:Female; D006801:Humans; D007371:Interferon-gamma / Q000187:drug effects / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000187:drug effects* / Q000378:metabolism; D008213:Lymphocyte Activation / Q000187:drug effects*; D008297:Male; D008875:Middle Aged; D010117:Oxypurinol / Q000009:adverse effects* / Q000494:pharmacology; D012016:Reference Values; D013262:Stevens-Johnson Syndrome / Q000209:etiology / Q000276:immunology; D013844:Thiazoles / Q000009:adverse effects / Q000494:pharmacology
chemical_list: D000945:Antigens, Differentiation, T-Lymphocyte; C075005:GNLY protein, human; D013844:Thiazoles; D000069465:Febuxostat; D000493:Allopurinol; D007371:Interferon-gamma; D010117:Oxypurinol