Graves病患者外周血CD4^+CD28^-T淋巴细胞的检测和分选

Detection and Sorting of CD4^+CD28^- T Lymphocytes in Peripheral Blood in Patients with Graves Disease

目的分析CD4^+CD28^-T淋巴细胞Graves病(毒性弥漫性甲状腺肿)患者外周血的表达及探索纯化该T细胞的有效方法。方法运用常规密度梯度法分离Graves病患者外周血单个核细胞,用CD4、CD28单克隆抗体进行免疫荧光双标记后进行流式细胞术检测和分选。结果Grave病患者外周血T细胞亚群中,CD4^+CD28^-的表达率明显高于正常人(P<0.01),采用流式细胞术和单克隆抗体双标记对CD4^+CD28^-T淋巴细胞进行快速无菌分选是可行的。结论流式细胞术可用于Graves病患者外周血CD4^+CD28^-T淋巴细胞的检测和分选。

Objective To detect CD4^＋CD28ˉ T cells in peripheral blood in patients with Graves disease（GD） and develop an effective measure of purifying theses T cells. Methods PBMCs（peripheral blood mononuclear cells） were isolated with routine density gradient centrifugation method, stained with fluorochrome conjugated antibodies, and analyzed and sorted by flow cytometry subsequently. Results It showed that the level of CD4^＋CD28ˉ T cells in peripheral blood T lymphocytes from GD patients was significantly higher than that from healthy subjects （P〈0.01）. And it was demonstrated to be an effective measure of sorting CD4^＋CD28ˉ T cells quickly and aseptically with two color flow cytometry mehtod. Conclusion Flow cytometry is suitable for detection and sorting of CD4^＋CD28ˉ T lymphocytes in peripheral blood of GD patients.