Oxypurinol-Specific T Cells Possess Preferential TCR Clonotypes and Express Granulysin in Allopurinol-Induced Severe Cutaneous Adverse Reactions
Oxypurinol-Specific T Cells Possess Preferential TCR Clonotypes and Express Granulysin in Allopurinol-Induced Severe Cutaneous Adverse Reactions
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DOI：
10.1038/jid.2015.165
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发表时间：
2015-09-01
影响因子：
6.5
通讯作者：
Hung, Shuen-Iu
中科院分区：
文献类型：
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作者：
Chung, Wen-Hung;Pan, Ren-You;Hung, Shuen-Iu
别嘌呤醇是治疗痛风和高尿酸血症的一线药物，是严重皮肤不良反应（scar）的主要原因之一。为了研究别嘌呤醇诱导SCAR的分子机制，我们招募了21例患者(13例Stevens-Johnson综合征（SJS)/中毒性表皮坏死松解（TEN）， 8例伴有嗜酸性粒细胞增多和全身症状的药物反应（DRESS））， 11例耐受对照和23例健康供体。我们用别嘌呤醇、氧嘌呤醇或非布司他培养外周血单个核细胞（PBMCs），并测量培养上清液中颗粒蛋白和ifn - γ的表达，进行了体外t细胞活化实验。采用新一代测序技术研究TCR曲目。氧嘌呤醇刺激导致疤痕患者（n = 14）血液样本培养物中颗粒蛋白显著增加，但耐受对照组（n = 11）或健康供者（n = 23）血液样本培养物中颗粒蛋白显著增加。氧嘌呤醇以浓度和时间依赖的方式诱导t细胞反应，而别嘌呤醇或非布司他则没有。别嘌呤醇- scar患者的T细胞与非布司他无交叉反应。在来自皮肤病变的水疱细胞（n = 8）和来自别嘌呤醇- scar患者的氧嘌呤醇活化t细胞培养（n = 4）中发现了tcr - v - β的优先使用和特异性CDR3（第三互补决定区）的克隆扩增。这些数据表明，除HLA-B*58:01外，在氧嘌呤醇诱导下表达颗粒蛋白的克隆型特异性T细胞参与了别嘌呤醇诱导的SCAR的发病机制。
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-gamma 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-beta 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.