钾通道阻滞剂对多发性硬化患者CD4^＋T细胞CCR7及CD45RA表达韵影响

Change of CCR7 and CD45RA after blocking of Kvl. 3 potassium channel of CD4 ^＋ T lymphocytes in multiple sclerosis

目的研究Kvl．3钾通道阻滞剂海葵毒素（SHK）阻滞多发性硬化（Ms）患者急性期髓鞘反应性CD4^＋T淋巴细胞上的Kvl．3钾通道后，细胞表型CCR7、CD45RA表达的变化与疾病的关系。方法对15例急性期MS患者、15例INF—β—1b治疗缓解期MS患者和15名健康对照的不同状态下的CD4^＋T淋巴细胞进行CD3、CD4、CCR7、CD45RA的荧光抗体及同型对照标记，用四标流式细胞仪检测3组不同状态下细胞表型CCR7、CD45RA表达的变化。结果Ms急性期组CD4^＋T细胞以CCR7-CD45RA^-（TEM）表型为主，其变化与病情有明显相关性，髓鞘抗原刺激后此型所占比例明显增高（P〈0．05），初始细胞表型CCR7^＋CD45RA^＋减少（P〈0．05），钾通道阻滞剂SHK对MS急性期患者髓鞘反应性CD4^＋TEM表型有明显抑制作用（P〈0．05）。结论检测CD4^＋TEM细胞表型可能对判断病情有一定临床意义，Kvl．3钾通道可能成为治疗MS的新靶点。

Objective To investigate the changes of CCR7 and CD45RA expression after blocking of the potassium channel Kvl. 3 in myelin specific CD4^ ＋ T lymphocytes and the relation thereof with multiple sclerosis（MS）. Methods Peripheral blood mononuclear cells were isolated from 15 activated MS patients, 15 INF-β-1b treated MS patients, and 15 normal controls, CD4^ ＋T lymphocytes were isolated using positive selection method with anti-CD4-coated magnetic beads. To establish culturing MBP special CD4^＋T lymphocyte lines, the different groups of T ceil were labeled with CD3, CD4, CCR7, and CD45RA fluorescence-antibody or homotype controls and analyzed by four-color flow cytometer. Results The most part of phenotype in the activated MS patients was CD4 ^＋ CCR7 - CD45RA- T cells and the percentage was increased after myelin antigen stimulation （ P 〈 0. 05 ）, whereas the percentage of CCR7 ^- CD45RA ^＋ T cells was decreased（P 〈0. 05）. SHK greatly inhibited CCR7 - CD45RA- in activated MS（P 〈0.05）. CCR7- CD45RA- and CCR7 ^＋ CD45RA- were in correlation with expanded disability status scale （EDSS） score （r = 0. 73 ,r = 0. 705 ,P 〈 0. 05 ） in the peripheral blood of activated MS. Conclusion There is a strong correlation between TEM phenotype and severity of MS, which may suggest Tem phenotype as the marker to estimate the state of illness. Kvl. 3 potassium channel may be the new target in treatment of MS.