Chu Tony L H, Guan Qiunong, Nguan Christopher Y C, Du Caigan
Department of Urologic Sciences, University of British Columbia, Vancouver, British Columbia, Canada.
Immunity and Infection Research Centre, Vancouver Coastal Health Research Institute, Vancouver, British Columbia, Canada.
PLoS One. 2015 Dec 15;10(12):e0144735. doi: 10.1371/journal.pone.0144735. eCollection 2015.
Both rapamycin (RAPA) and cyclosporin A (CsA) are commonly used for immunosuppression, however their adverse side effects limit their application. Thus, it is of interest to develop novel means to enhance or preserve the immunosuppressive activity of RAPA or CsA while reducing their toxicity. Halofuginone (HF) has been recently tested as a potential immunosuppressant. This study investigated the interaction of HF with RAPA or with CsA in cell cultures. Cell proliferation in cultures was determined using methylthiazol tetrazolium assay, and cell apoptosis assessed by flow cytometric analysis and Western blot. The drug-drug interaction was determined according to Loewe's equation or Bliss independence. Here, we showed that addition of HF to anti-CD 3 antibody-stimulated splenocyte cultures induced synergistic suppression of T cell proliferation in the presence of RAPA, indicated by an interaction index (γ) value of < 1.0 between HF and RAPA, but not in those with CsA. The synergistic interaction of RAPA with HF in the suppression of T cell proliferation was also seen in a mixed lymphocyte reaction and Jurkat T cell growth, and was positively correlated with an increase in cell apoptosis, but not with proline depletion. In cultured kidney tubular epithelial cells, HF attenuated the cytotoxicity of CsA. In conclusion, these data indicate that HF synergistically enhances anti-T cell proliferation of RAPA and reduces the nephrotoxicity of CsA in vitro, suggesting the potential use of HF for enhancing anti-T cell proliferation of RAPA and reducing CsA-mediated nephrotoxicity.
雷帕霉素（RAPA）和环孢素A（CsA）都常用于免疫抑制，但它们的副作用限制了其应用。因此，开发新方法来增强或保留RAPA或CsA的免疫抑制活性同时降低其毒性具有重要意义。卤夫酮（HF）最近已作为一种潜在的免疫抑制剂进行了测试。本研究在细胞培养中探究了HF与RAPA或与CsA的相互作用。使用甲基噻唑四氮唑法测定培养物中的细胞增殖，并通过流式细胞术分析和蛋白质印迹法评估细胞凋亡。根据洛伊方程或布利斯独立性来确定药物 - 药物相互作用。在此，我们表明在抗CD 3抗体刺激的脾细胞培养物中添加HF会在存在RAPA的情况下诱导对T细胞增殖的协同抑制，HF与RAPA之间的相互作用指数（γ）值<1.0表明了这一点，但在添加CsA的培养物中则没有。RAPA与HF在抑制T细胞增殖方面的协同相互作用在混合淋巴细胞反应和Jurkat T细胞生长中也可见，并且与细胞凋亡的增加呈正相关，但与脯氨酸消耗无关。在培养的肾小管上皮细胞中，HF减轻了CsA的细胞毒性。总之，这些数据表明HF在体外协同增强RAPA的抗T细胞增殖作用并降低CsA的肾毒性，提示HF在增强RAPA的抗T细胞增殖作用和降低CsA介导的肾毒性方面具有潜在用途。
