Laboratory of Toxicology, Dipartimento di Scienze e Politiche Ambientali, Università degli Studi di Milano, Via Balzaretti 9, 20133 Milano, Italy.
Laboratory of Toxicology, Dipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, Via Balzaretti 9, 20133 Milano, Italy.
Toxicol In Vitro. 2020 Oct;68:104953. doi: 10.1016/j.tiv.2020.104953. Epub 2020 Jul 28.
Hypersensitivity drug reactions (HDRs) are common among drugs, despite this, there are no validated in vitro or in vivo methods for screening the sensitizing potential of drugs in the preclinical phase. We previously developed the THP-1 activation assay, based on CD86 upregulation and IL-8 production, for the in vitro identification of drugs able to induce selective dendritic cell activation. In this paper, we investigated the predictive capacity of the method toward drugs associated with HDRs for which a correlation with specific human leukocyte antigens (HLA) have been demonstrated. For that purpose, abacavir, carbamazepine and clozapine were used. Metformin was used as negative control. Dose- and time-course experiments were conducted. The surface markers CD86, CD54 and HLA-DR were evaluated by flow cytometry analysis, whereas IL-8 release by ELISA. Abacavir, carbamazepine and clozapine gave positive results with CD86 upregulation and/or IL-8 release, with abacavir also inducing HLA-DR. The test reveals the ability of drugs to induce dendritic cell activation (signals 1/2), that preceded the adaptive immune response, which will be manifested only in a minority of patients carrying the specific HLA genotypes. The idea is to integrate this simple method during drug development to identify the potential of drugs to induce hypersensitivity reactions in the pre-clinical phase.
药物超敏反应（HDRs）在药物中很常见，尽管如此，目前还没有经过验证的体外或体内方法可用于在临床前阶段筛选药物的致敏潜力。我们之前开发了基于 CD86 上调和 IL-8 产生的 THP-1 激活测定法，用于体外鉴定能够诱导选择性树突状细胞激活的药物。在本文中，我们研究了该方法对与 HDR 相关的药物的预测能力，这些药物与特定的人类白细胞抗原（HLA）相关联已得到证实。为此，使用了阿巴卡韦、卡马西平和氯氮平。二甲双胍用作阴性对照。进行了剂量和时间过程实验。通过流式细胞术分析评估表面标记物 CD86、CD54 和 HLA-DR，通过 ELISA 评估 IL-8 释放。阿巴卡韦、卡马西平和氯氮平均通过 CD86 上调和/或 IL-8 释放产生阳性结果，阿巴卡韦还诱导 HLA-DR。该测试揭示了药物诱导树突状细胞激活的能力（信号 1/2），这先于适应性免疫反应，而只有携带特定 HLA 基因型的少数患者才会出现这种反应。其想法是在药物开发过程中整合这种简单的方法，以在临床前阶段识别药物引起过敏反应的潜力。
