Characterization of the formation and localization of sulfamethoxazole and dapsone-associated drug-protein adducts in human epidermal keratinocytes
Characterization of the formation and localization of sulfamethoxazole and dapsone-associated drug-protein adducts in human epidermal keratinocytes
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DOI：
10.1124/jpet.105.086009
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发表时间：
2005-07-01
影响因子：
3.5
通讯作者：
Svensson, CK
中科院分区：
文献类型：
--
作者：
Roychowdhury, S;Vyas, PM;Svensson, CK
磺胺和砜诱导的超敏反应被认为是通过母药分子(S)对各自的反应性代谢物(S)的生物激活而介导的。最近的研究表明，角质形成细胞可以激活磺胺类和砜类化合物。应用酶联免疫吸附试验和本实验室研制的兔半抗原特异性抗血清，发现正常人表皮角质形成细胞(NHEKs)或永生化角质形成细胞系(HaCaT)与磺胺甲恶唑(SMX)或氨苯砜(DDS)孵育均可形成药物/代谢物蛋白加合物。在抗坏血酸存在下，这些与SMX的加合物的形成增加，而N-乙酰半胱氨酸减少与SMX和DDS的加合物的形成。当NHEK与SMX、DDS或它们各自的芳羟胺代谢物孵育时，共聚焦显微镜证实了加合物的形成。比较了渗透性和非渗透性NHEK中加合物的细胞分布。暴露于SMX、DDS或氨苯松羟胺可导致细胞内加合物的形成，而SMX羟胺也可导致细胞表面加合物的存在。综上所述，我们的工作表明，角质形成细胞可以激活SMX/DDS形成药物-蛋白质加合物，该加合物可能在角质形成细胞死亡时由抗原提呈细胞获得，从而引发免疫反应。此外，角质形成细胞本身也可能向半抗原特异性细胞毒性T淋巴细胞递呈抗原。此外，我们的结果还表明，不同的磺胺类/磺酸类化合物在角质形成细胞中可能有不同的原位触觉蛋白靶点。
Sulfonamide- and sulfone- induced hypersensitivity reactions are thought to be mediated through bioactivation of parent drug molecule( s) to their respective reactive metabolite( s). Recent studies have demonstrated that keratinocytes can bioactivate sulfonamides and sulfones. Using enzyme- linked immunosorbent assay and hapten- specific rabbit antisera developed in our laboratory, we found that incubation of either normal human epidermal keratinocytes ( NHEKs) or an immortalized human keratinocyte cell line ( HaCaT) with sulfamethoxazole ( SMX) or dapsone ( DDS) resulted in the formation of drug/ metabolite protein adducts. The formation of these adducts with SMX was increased in the presence of ascorbic acid, whereas N- acetylcysteine decreased adduct formation with both SMX and DDS. Adduct formation was confirmed using confocal microscopy when NHEKs were incubated with SMX, DDS, or their respective arylhydroxylamine metabolites. Cellular distribution of adducts was compared in permeable versus nonpermeable NHEKs. Exposure to SMX, DDS, or dapsone hydroxylamine resulted in the formation of intracellular adducts, whereas SMX hydroxylamine also resulted in the presence of adducts on the cell surface. In summary, our work shows that keratinocytes can bioactivate SMX/ DDS to form drug- protein adducts, which may be acquired by antigen- presenting cells upon keratinocyte cell death, evoking an immune response. In addition, keratinocytes may themselves present antigen to hapten- specific cytotoxic T lymphocytes. Furthermore, our results also suggest that different sulfonamides/ sulfones may have different protein targets for in situ haptenation in keratinocytes.