Department of Medical Sciences School of Veterinary Medicine University of Wisconsin-Madison Madison WI USA.
Present address: Department of Veterinary Clinical Medicine College of Veterinary Medicine University of Illinois Urbana IL USA.
Pharmacol Res Perspect. 2018 Mar 2;6(2):e00388. doi: 10.1002/prp2.388. eCollection 2018 Apr.
The lymphocyte toxicity assay (LTA) is a proposed surrogate marker of sulfonamide antibiotic hypersensitivity. In the LTA, peripheral blood mononuclear cells (PBMCs) undergo apoptosis more readily in hypersensitive versus tolerant patients when exposed to drug-hydroxylamine metabolites in vitro. The purpose of this study was to identify key gene transcripts associated with increased cytotoxicity from sulfamethoxazole-hydroxylamine in human PBMCs in the LTA. The LTA was performed on PBMCs of 10 patients hypersensitive to trimethoprim-sulfamethoxazole (HS) and 10 drug-tolerant controls (TOL), using two cytotoxicity assays: YO-PRO (n = 20) and MTT (n = 12). mRNA expression profiles of PBMCs, enriched for CD8 T cells, were compared between HS and TOL patients. Transcript expression was interrogated for correlation with % cytotoxicity from YO-PRO and MTT assays. Correlated transcripts of interest were validated by qPCR. LTA results were not significantly different between HS and TOL patients, and no transcripts were found to be differentially expressed between the two groups. 96 transcripts were correlated with cytotoxicity by YO-PRO ( = ±.63-.75, FDR 0.188). Transcripts were selected for validation based on mechanistic plausibility and three were significantly over-expressed by qPCR in high cytotoxicity patients: multi-specific organic anion transporter C (), mitoferrin-1 (), and Porimin (). These data identify novel transcripts that could contribute to sulfonamide-hydroxylamine induced cytotoxicity. These include , encoding a mitochondrial iron transporter, , encoding an arylamine drug transporter, and , encoding a transmembrane protein that mediates cell death.
淋巴细胞毒性测定（LTA）是一种磺胺类抗生素过敏的替代标志物。在 LTA 中，当体外接触药物羟胺代谢物时，与耐受患者相比，敏感患者的外周血单核细胞（PBMC）更容易发生细胞凋亡。本研究的目的是鉴定与 LTA 中磺胺甲恶唑羟胺导致的人 PBMC 细胞毒性增加相关的关键基因转录本。在 10 例对甲氧苄啶磺胺甲恶唑（HS）过敏的患者（HS）和 10 例药物耐受对照（TOL）中进行了 LTA，使用了两种细胞毒性测定法：YO-PRO（n=20）和 MTT（n=12）。比较了 HS 和 TOL 患者 PBMC 的 CD8 T 细胞富集的 mRNA 表达谱。对 PBMC 转录本表达进行了分析，以了解其与 YO-PRO 和 MTT 测定的细胞毒性的相关性。对感兴趣的相关转录本进行了 qPCR 验证。HS 和 TOL 患者的 LTA 结果无显著差异，两组之间未发现差异表达的转录本。有 96 个转录本与 YO-PRO 的细胞毒性相关（=±.63-.75，FDR 0.188）。根据机制的合理性选择了验证的转录本，并且在高细胞毒性患者中，qPCR 验证了三个转录本显著过表达：多特异性有机阴离子转运蛋白 C（）、线粒体铁转运蛋白 1（）和 Porimin（）。这些数据确定了可能导致磺胺类/羟胺诱导的细胞毒性的新转录本。这些包括编码线粒体铁转运蛋白的，编码芳基胺药物转运蛋白的，编码介导细胞死亡的跨膜蛋白的。
