软木酰苯胺氧肟酸免疫调节功能研究

Evaluation of Immunomodulation by Histone Deacetylase Inhibitor SAHA

目的通过体外实验观察软木酰苯胺氧肟酸(SAHA)对效应T细胞和调节性T细胞的作用。方法①淋巴细胞增殖的检测:取CFSE标记的淋巴细胞、CD4+或CD8+T细胞作为反应细胞,实验组加入不同浓度SAHA及CD3/CD28单抗作为刺激原培养,设阴性对照组(仅加入CD3/CD28单抗)和空白对照组,96 h后检测各组细胞增殖情况;②以淋巴细胞作为反应细胞,实验组中加入不同浓度SAHA及CD3/CD28单抗作为刺激原培养,设阴性对照组(仅加入CD3/CD28单抗)和空白对照组。96 h后检测各组CD4+Foxp3+T细胞比例。结果①SAHA剂量依赖性抑制CD3/CD28单抗诱导的淋巴细胞、CD4+和CD8+T细胞增殖(均P<0.05);②CD3/CD28单抗不能诱导CD4+Foxp3+T细胞比例增高,1μmol.L-1SAHA也未能诱导CD4+Foxp3+T细胞比例上调。但给予3和5μmol.L-1SAHA后,CD4+Foxp3+T细胞比例显著提高,组内和组间比较,均差异有显著性(均P<0.05)。结论体外实验表明,SAHA具有免疫调节功能,不仅能抑制效应性T细胞增殖,而且一定浓度药物作用能够促使调节性T细胞比例上调。

ObjectiveTo investigate effects of histone deacetylase inhibitor SAHA on effector and regulatory T cells in vitro.Methods①As the reactive cells, lymphocyte, CD＋4 T and CD＋8 T cells labelled with CFSE were stimulated with anti-CD3 and anti-CD28 mAbs in the presence and absence of SAHA （experimental group and control group） at different level, or no any stimulus instead of PBS （placebo group）. After 96 hours, the proliferation of cells in each group was detected. ②As the reactive cells, lymphocyte were stimulated with anti-CD3 mAb and anti-CD28 mAb in the presence and absence of SAHA （experimental group and control group）, or no any stimulus instead of PBS （placebo group）. After 96 hours, the percentage changes of CD＋4Foxp3＋ T cells in each group were detected. Results①The proliferation of CFSE-labelled lymphocyte, CD＋4 T and CD＋8 T cells mediated with anti-CD3 and anti-CD28 mAbs were inhibited by SAHA in a dose dependent manner（P〈0.05）. ②In contrast to placebo group, anti-CD3 and anti-CD28 mAbs and SAHA at a final concentration of 1 μmol＆#8226;L-1 stimulation did not enhance the percentage of CD＋4Foxp3＋ T cells（P〉0.05）. But in combination with SAHA at final concentration of 3 μmol＆#8226;L-1 and 5 μmol＆#8226;L-1, the percentage of CD＋4Foxp3＋ T cells was notably elevated（P〈0.05）. ConclusionOur data suggest that SAHA has immunomodulation effects, which not only directly inhibiting T-effector cells proliferation but also increasing Treg cells with certain concentration in vitro.