Faulkner Lee, Gibson Andrew, Sullivan Andrew, Tailor Arun, Usui Toru, Alfirevic Ana, Pirmohamed Munir, Naisbitt Dean J, Kevin Park B
Department of Molecular and Clinical Pharmacology, Centre for Drug Safety Science;
Department of Personalised Medicine, Institute of Translational Medicine, University of Liverpool, Liverpool, UK
Toxicol Sci. 2016 Dec;154(2):416-429. doi: 10.1093/toxsci/kfw177. Epub 2016 Sep 16.
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-B15:02 donors), flucloxacillin (in 1 HLA-B57:01 donor) and oxypurinol (in HLA-B58:01 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-B57:01 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.
许多严重的药物不良反应是由T细胞引起的。已确定某些反应与HLA等位基因有关，这使得开发标准化的临床前试验来预测化学毒性变得困难。我们最近开发了一种使用药物代谢物亚硝基磺胺甲恶唑（SMX-NO）的T细胞致敏试验。我们现在报告该试验的可重复性、从1000名HLA分型志愿者建立外周血单核细胞生物样本库以及对一组化合物产生抗原特异性反应的情况。用SMX-NO进行了40次T细胞致敏试验；5次反应较弱（1.5-1.9），34次显示良好（SI 2.0-3.9）或强烈反应（SI＞4.0），采用增殖和细胞因子释放的读数。因此，SMX-NO可作为体外T细胞活化的模型试剂。对与HLA风险等位基因无关的半抗原化合物（阿莫西林、哌拉西林和班德罗夫斯基碱）也产生了良好到强烈的反应。此外，在与HLA-I类限制性超敏反应相关的卡马西平（在HLA-B15:02供体中）、氟氯西林（在1名HLA-B57:01供体中）和氧嘌呤醇（在HLA-B58:01供体中）中检测到反应。相比之下，对ximelagatran、氯美昔布和拉帕替尼（HLA-II类限制性超敏反应形式）的幼稚T细胞致敏产生了阴性结果。阿巴卡韦可激活患者的记忆T细胞，但不能激活来自HLA-B57:01供体的幼稚T细胞。这项工作表明，致敏试验可用于评估T细胞对药物的初始反应，并研究显示HLA-I类限制性的药物的T细胞致敏机制。需要进一步的研究来调查HLA-II类限制性反应。
