RIKILT Institute of Food Safety, Wageningen University and Research Centre, P.O. Box 230, 6700 AE, Wageningen, The Netherlands.
Department of Toxicogenomics, Maastricht University, P.O. Box 616, 6200 MD, Maastricht, The Netherlands.
Arch Toxicol. 2015 Dec;89(12):2325-37. doi: 10.1007/s00204-014-1365-9. Epub 2014 Sep 17.
Transcriptomics in combination with in vitro cell systems is a powerful approach to unravel modes of action of toxicants. An important question is to which extent the modes of action as revealed by transcriptomics depend on cell type, species and study type (in vitro or in vivo). To acquire more insight into this, we assessed the transcriptomic effects of the immunosuppressive drug cyclosporine A (CsA) upon 6 h of exposure of the mouse cytotoxic T cell line CTLL-2, the thymoma EL-4 and primary splenocytes and compared these to the effects in spleens of mice orally treated with CsA for 7 days. EL-4 and CTLL-2 cells showed the highest similarities in response. CsA affected many genes in primary splenocytes that were not affected in EL-4 or CTLL-2. Pathway analysis demonstrated that CsA upregulated the unfolded protein response, endoplasmic reticulum stress and NRF2 activation in EL-4 cells, CTLL-2 cells and primary mouse splenocytes but not in mouse spleen in vivo. As expected, CsA downregulated cell cycle and immune response in splenocytes in vitro, spleens in vivo as well as CTLL-2 in vitro. Genes up- and downregulated in human Jurkat, HepG2 and renal proximal tubular cells were similarly affected in CTLL-2, EL-4 and primary splenocytes in vitro. In conclusion, of the models tested in this study, the known mechanism of immunotoxicity of CsA is best represented in the mouse cytotoxic T cell line CTLL-2. This is likely due to the fact that this cell line is cultured in the presence of a T cell activation stimulant (IL-2) making it more suitable to detect inhibitory effects on T cell activation.
转录组学与体外细胞系统相结合是揭示毒物作用模式的有力方法。一个重要的问题是，转录组学揭示的作用模式在多大程度上取决于细胞类型、物种和研究类型（体外或体内）。为了更深入地了解这一点，我们评估了免疫抑制剂环孢素 A（CsA）在暴露于小鼠细胞毒性 T 细胞系 CTLL-2、胸腺瘤 EL-4 和原代脾细胞 6 小时后的转录组效应，并将其与 CsA 经口处理 7 天的小鼠脾脏效应进行了比较。EL-4 和 CTLL-2 细胞在反应上表现出最高的相似性。CsA 影响了许多在 EL-4 或 CTLL-2 中不受影响的原代脾细胞中的基因。通路分析表明，CsA 上调了 EL-4 细胞、CTLL-2 细胞和原代小鼠脾细胞中的未折叠蛋白反应、内质网应激和 NRF2 激活，但在体内小鼠脾脏中没有。正如预期的那样，CsA 下调了体外脾细胞、体内脾脏以及 CTLL-2 中的细胞周期和免疫反应。在体外 Jurkat、HepG2 和肾近端肾小管细胞中上调和下调的基因在 CTLL-2、EL-4 和原代脾细胞中也受到类似的影响。总之，在本研究中测试的模型中，CsA 的已知免疫毒性机制在小鼠细胞毒性 T 细胞系 CTLL-2 中得到了最好的体现。这可能是因为该细胞系在 T 细胞激活刺激物（IL-2）的存在下培养，使其更适合检测对 T 细胞激活的抑制作用。
