Institute for Tissue Transplantation and Immunology, Jinan University, Guangzhou, Guangdong, 510632, China.
Inflamm Res. 2012 Jun;61(6):623-34. doi: 10.1007/s00011-012-0454-6. Epub 2012 Mar 10.
FTY720 is a potent drug for multiple sclerosis treatment. To biologically address its possible applications to more generalized diseases with aberrant inflammation, we are testing whether FTY720 can function as a lymphocyte cell cycle blocker and activation suppressor via a concanavalin A (ConA)-mediated mouse lymphocyte pan-activation model.
Mouse lymphocytes were obtained from lymph nodes and subjected to ConA and/or FTY720 treatment. Cell viability was assayed by MTT and mitochondrial assays. Early and late activation, cell cycle, proliferation and intracellular Ca(2+) concentration (Ca(2+)) were analyzed by flow cytometry.
At concentrations of less than 500 nM, FTY720 significantly down-regulated both CD69 and CD25 expressions of T cells, as well as inhibiting proliferation of activated lymphocytes. In addition, FTY720 blocked the ConA-induced mitogenesis, exhibiting lymphocyte G(0)/G(1) phase cell cycle arrest with significant reduction of cells in S and G(2)/M phases. Meanwhile, a significant decline in Ca(2+) was observed. The correlation of Ca(2+) and cell cycle arrest were validated by employing a Ca(2+) inhibitor SK&F 96365 and testing with and without FTY720 treatment.
We demonstrated that FTY720 induces G(0)/G(1) phase cell cycle arrest, resulting in proliferation inhibition upon lymphocyte pan-activation, which may be related to reduction of overall intracellular Ca(2+) load.
FTY720 是一种有效的多发性硬化症治疗药物。为了从生物学角度研究其在炎症异常的更广泛疾病中的潜在应用，我们正在测试 FTY720 是否可以通过伴刀豆球蛋白 A（ConA）介导的小鼠淋巴细胞全激活模型作为淋巴细胞细胞周期阻断剂和激活抑制剂发挥作用。
从小鼠淋巴结中获得淋巴细胞，并进行 ConA 和/或 FTY720 处理。通过 MTT 和线粒体测定法测定细胞活力。通过流式细胞术分析早期和晚期激活、细胞周期、增殖和细胞内 Ca（2+）浓度([Ca（2+）]（i）)。
在浓度低于 500 nM 的情况下，FTY720 显著下调 T 细胞的 CD69 和 CD25 表达，并抑制活化淋巴细胞的增殖。此外，FTY720 阻断了 ConA 诱导的有丝分裂，表现为淋巴细胞 G0/G1 期细胞周期阻滞，S 期和 G2/M 期细胞显著减少。同时，观察到 [Ca（2+）]（i）显著下降。通过使用 Ca（2+）抑制剂 SK&F 96365 并在有无 FTY720 处理的情况下进行测试，验证了 [Ca（2+）]（i）与细胞周期阻滞的相关性。
我们证明 FTY720 诱导 G0/G1 期细胞周期阻滞，导致淋巴细胞全激活时增殖抑制，这可能与整体细胞内 Ca（2+）负荷减少有关。
