Dapsone (DDS) causes hypersensitivity reactions in 0.5–3.6% of patients. Although clinical diagnosis is indicative of a hypersensitivity reaction, studies have not been performed to define whether dapsone or a metabolite activates specific T-cells. Thus, the aims of this study were to explore the immunogenicity DDS and nitroso DDS (DDS-NO) using peripheral blood mononuclear cells from healthy donors and splenocytes from mice and generate human T-cell clones to characterize mechanisms of T-cell activation. DDS-NO was synthesized from DDS-hydroxylamine and shown to bind to the thiol group of glutathione and human and mouse albumin through sulfonamide and N-hydroxyl sulphonamide adducts. Naïve T-cell priming to DDS and DDS-NO was successful in three human donors. DDS-specific CD4+ T-cell clones were stimulated to proliferate in response to drug via a MHC class II restricted direct binding interaction. Cross reactivity with DDS-NO, DDS-analogues, and sulfonamides was not observed. DDS-NO clones were CD4+ and CD8+, MHC class II and I restricted, respectively, and activated via a pathway dependent on covalent binding and antigen processing. DDS and DDS-NO-specific clones secreted a mixture of Th1 and Th2 cytokines, but not granzyme-B. Splenocytes from mice immunized with DDS-NO were stimulated to proliferate in vitro with the nitroso metabolite, but not DDS. In contrast, immunization with DDS did not activate T-cells. These data show that DDS- and DDS-NO-specific T-cell responses are readily detectable.

中文翻译：

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氨苯砜和亚硝基氨苯砜激活来自健康供体的幼稚 T 细胞

氨苯砜 (DDS) 在 0.5-3.6% 的患者中引起超敏反应。尽管临床诊断表明过敏反应，但尚未进行研究以确定氨苯砜或代谢物是否激活特定的 T 细胞。因此，本研究的目的是利用来自健康供体的外周血单核细胞和来自小鼠的脾细胞探索免疫原性 DDS 和亚硝基 DDS (DDS-NO)，并生成人类 T 细胞克隆来表征 T 细胞活化的机制。DDS-NO 由 DDS-羟胺合成，并显示通过磺胺和N与谷胱甘肽和人和小鼠白蛋白的硫醇基团结合-羟基磺酰胺加合物。对 DDS 和 DDS-NO 的初始 T 细胞启动在三个人类供体中是成功的。通过 MHC II 类限制的直接结合相互作用，刺激 DDS 特异性 CD4+ T 细胞克隆增殖以响应药物。未观察到与 DDS-NO、DDS 类似物和磺胺类药物的交叉反应。DDS-NO 克隆分别受到 CD4+ 和 CD8+、MHC II 类和 I 类限制，并通过依赖于共价结合和抗原加工的途径激活。DDS 和 DDS-NO 特异性克隆分泌 Th1 和 Th2 细胞因子的混合物，但不分泌颗粒酶 B。来自用 DDS-NO 免疫的小鼠的脾细胞在体外被刺激增殖与亚硝基代谢物，但不是 DDS。相比之下，用 DDS 免疫不会激活 T 细胞。这些数据表明 DDS 和 DDS-NO 特异性 T 细胞反应很容易检测到。