Kim B S, Kim J Y, Lee J G, Cho Y, Huh K H, Kim M S, Kim Y S
The Research Institute for Transplantation, Yonsei University College of Medicine, Seoul, Republic of Korea; Department of Internal Medicine Severance Hospital, Yonsei University Health System, Seoul, Republic of Korea.
The Research Institute for Transplantation, Yonsei University College of Medicine, Seoul, Republic of Korea.
Transplant Proc. 2015 Apr;47(3):787-90. doi: 10.1016/j.transproceed.2014.12.038.
Thalidomide was originally used to alleviate morning sickness in pregnant women, but was banned due to severe adverse effects. Since the discovery of its anticancer and anti-inflammatory properties, it has regained research interest. However, its mechanism of action is still unknown. Therefore, we examined the effects of thalidomide on effector T (Teff) and regulatory T (Treg) cells in splenocytes of mice.
Splenic CD4(+), CD44(low), and CD62L(high) T lymphocytes (Tnaives) isolated from C57BL/6 mice were cultured for T-cell proliferation and Treg conversion. For T-cell proliferation, naive T cells (Tnaives) were cultured for 72 hours with anti-CD3 and anti-CD28 antibodies, and carboxyfluorescein succinimidyl ester (CFSE) labeling method was used. For Treg conversion, Tnaives were cultured for 72 hours with transforming growth factor-β1 (TGF-β1) and interleukin-2 (IL-2). Naïve T cells were plated at 1.5 × 10(5) cells on 96-well plates with 0, 1, 10, 50, or 100 μmol/L thalidomide. All samples were analyzed by flow cytometry after staining with CFSE, APC-conjugated anti-mouse CD4, and FITC-conjugated anti-mouse FoxP3.
Thalidomide significantly decreased the proliferation of CD4(+) Teffs in a dose-dependent manner (P < .01). In contrast, conversion to CD4(+)FoxP3(+) Tregs tended to increase by thalidomide treatment, although the increase was not statistically significant.
These findings suggest that thalidomide may have an immune modulatory effect by selectively suppressing CD4(+) Teff proliferation. Further studies will be needed to elucidate the underlying signaling pathway.
沙利度胺最初用于缓解孕妇的晨吐症状，但因其严重的不良反应而被禁用。自发现其抗癌和抗炎特性以来，它重新引起了研究兴趣。然而，其作用机制仍不清楚。因此，我们研究了沙利度胺对小鼠脾细胞中效应T（Teff）细胞和调节性T（Treg）细胞的影响。
从C57BL/6小鼠中分离出脾CD4(+)、CD44(low)和CD62L(high) T淋巴细胞（初始T细胞），用于T细胞增殖和Treg转化培养。对于T细胞增殖，初始T细胞（Tnaives）与抗CD3和抗CD28抗体一起培养72小时，并使用羧基荧光素琥珀酰亚胺酯（CFSE）标记法。对于Treg转化，Tnaives与转化生长因子-β1（TGF-β1）和白细胞介素-2（IL-2）一起培养72小时。将初始T细胞以1.5×10(5)个细胞接种到含有0、1、10、50或100μmol/L沙利度胺的96孔板中。在用CFSE、APC偶联的抗小鼠CD4和FITC偶联的抗小鼠FoxP3染色后，所有样品通过流式细胞术进行分析。
沙利度胺以剂量依赖性方式显著降低CD4(+) Teffs的增殖（P <.01）。相比之下，沙利度胺治疗虽使转化为CD4(+)FoxP3(+) Tregs的比例有增加趋势，但增加无统计学意义。
这些发现表明沙利度胺可能通过选择性抑制CD4(+) Teff增殖而具有免疫调节作用。需要进一步研究以阐明潜在的信号通路。
