Detection of Hepatic Drug Metabolite-Specific T-Cell Responses Using a Human Hepatocyte, Immune Cell Coculture System.
Detection of Hepatic Drug Metabolite-Specific T-Cell Responses Using a Human Hepatocyte, Immune Cell Coculture System.
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DOI：
10.1021/acs.chemrestox.2c00343
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发表时间：
2023-03-20
影响因子：
4.1
通讯作者：
Naisbitt, Dean John
中科院分区：
文献类型：
--
作者：
Ali, Serat-E;Meng, Xiaoli;Kafu, Laila;Hammond, Sean;Zhao, Qing;Ogese, Monday;Sison-Young, Rowena;Jones, Robert;Chan, Benjamin;Livoti, Lucia;Sun, Yonghu;Sun, Lele;Liu, Hong;Topping, Anthony;Goldring, Christopher;Zhang, Furen;Naisbitt, Dean John
药物应答性T细胞通常通过不同的途径（药理学相互作用和半抗原）被母体化合物或代谢物激活。研究药物超敏反应的一个障碍是缺乏用于功能研究的活性代谢物和缺乏原位产生代谢物的共培养系统。因此，本研究的目的是利用来自过敏患者的氨苯砜代谢物应答性T细胞，以及原代人肝细胞来驱动代谢物形成和随后的药物特异性T细胞应答。亚硝基氨苯砜反应性T细胞克隆是从过敏患者中产生的，其特征在于交叉反应性和T细胞活化途径。原代人肝细胞、抗原呈递细胞和T细胞共培养物以各种形式建立，其中肝脏和免疫细胞分离以避免细胞接触。将培养物暴露于氨苯砜，并分别通过LC-MS和增殖评估测量代谢物形成和T细胞活化。研究发现，当暴露于药物代谢物时，来自过敏患者的亚硝基氨苯砜响应性CD 4 + T细胞克隆以剂量依赖性方式增殖和分泌细胞因子。用亚硝基氨苯砜脉冲的抗原呈递细胞激活克隆，而固定抗原呈递细胞或从测定中省略抗原呈递细胞废除亚硝基氨苯砜特异性T细胞应答。重要的是，克隆与母体药物没有交叉反应性。在肝细胞免疫细胞共培养物的上清液中检测到亚硝基氨苯砜谷胱甘肽结合物，表明肝细胞衍生的代谢产物形成并转移到免疫细胞区室。类似地，亚硝基氨苯砜响应克隆被刺激增殖与氨苯砜，当肝细胞被添加到共培养系统。总的来说，我们的研究表明使用肝细胞免疫细胞共培养系统来检测原位代谢物形成和代谢物特异性T细胞反应。在未来的诊断和预测测定中应使用类似的系统，以在无法获得合成代谢物时检测代谢物特异性T细胞反应。
Drug-responsive T-cells are activated with the parent compound or metabolites, often via different pathways (pharmacological interaction and hapten). An obstacle to the investigation of drug hypersensitivity is the scarcity of reactive metabolites for functional studies and the absence of coculture systems to generate metabolites in situ. Thus, the aim of this study was to utilize dapsone metabolite-responsive T-cells from hypersensitive patients, alongside primary human hepatocytes to drive metabolite formation, and subsequent drug-specific T-cell responses. Nitroso dapsone-responsive T-cell clones were generated from hypersensitive patients and characterized in terms of cross-reactivity and pathways of T-cell activation. Primary human hepatocytes, antigen-presenting cells, and T-cell cocultures were established in various formats with the liver and immune cells separated to avoid cell contact. Cultures were exposed to dapsone, and metabolite formation and T-cell activation were measured by LC–MS and proliferation assessment, respectively. Nitroso dapsone-responsive CD4+ T-cell clones from hypersensitive patients were found to proliferate and secrete cytokines in a dose-dependent manner when exposed to the drug metabolite. Clones were activated with nitroso dapsone-pulsed antigen-presenting cells, while fixation of antigen-presenting cells or omission of antigen-presenting cells from the assay abrogated the nitroso dapsone-specific T-cell response. Importantly, clones displayed no cross-reactivity with the parent drug. Nitroso dapsone glutathione conjugates were detected in the supernatant of hepatocyte immune cell cocultures, indicating that hepatocyte-derived metabolites are formed and transferred to the immune cell compartment. Similarly, nitroso dapsone-responsive clones were stimulated to proliferate with dapsone, when hepatocytes were added to the coculture system. Collectively, our study demonstrates the use of hepatocyte immune cell coculture systems to detect in situ metabolite formation and metabolite-specific T-cell responses. Similar systems should be used in future diagnostic and predictive assays to detect metabolite-specific T-cell responses when synthetic metabolites are not available.
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DOI：
10.1111/j.1365-2125.1995.tb05797.x
发表时间：
1995-12-01
影响因子：
3.4
作者：
Gill, HJ;Tingle, MD;Park, BK
通讯作者：
Park, BK
DOI：
10.1124/jpet.105.086009
发表时间：
2005-07-01
影响因子：
3.5
作者：
Roychowdhury, S;Vyas, PM;Svensson, CK
通讯作者：
Svensson, CK
影响因子：
3.8
作者：
Tailor, Arun;Meng, Xiaoli;Naisbitt, Dean J.
通讯作者：
Naisbitt, Dean J.
影响因子：
15.9
作者：
Zanni, MP;von Greyerz, S;Pichler, WJ
通讯作者：
Pichler, WJ
影响因子：
3.8
作者：
Faulkner, Lee;Martinsson, Klara;Park, B. Kevin
通讯作者：
Park, B. Kevin