CD4^＋CD25^-Foxp3^＋T淋巴细胞的表型鉴定及其在初发系统性红斑狼疮中的意义

Phenotypfc difference between CD4^＋CD25^-Foxp3^＋ and CD4^＋CD25^＋Foxp3^＋ T cells and its clinical relevance in patients with untreated new-onset lupus

目的对初发系统性红斑狼疮（SLE）患者外周血异常表达CD4^＋CD25^-Foxp3^＋T淋巴细胞进行表型鉴定，并探讨其临床意义。方法对初发SLE患者外周血CD4^＋T淋巴细胞进行细胞表面分子[CD25、CD127、CCR4、糖皮质激素诱导的肿瘤坏死因子受体（GITR）、细胞毒T淋巴细胞相关抗原4（CT-LA-4）]和胞内分子（Foxp3）标染，流式细胞仪检测，并研究CD4^＋各细胞亚群与狼疮肾炎和疾病活动度（SLEDAI）相关性。结果SLE患者外周血CD4^＋CD25^-Foxp3^＋T淋巴细胞表面GITR、CTLA-4和CCR4表达率与活化T淋巴细胞（CD4^＋CD25^＋Foxp3^-）相比差异无统计学意义（P均〉0．05），而显著低于调节性T淋巴细胞（CD4^＋CD25^＋Foxp3^＋）（P均〈0．01）；CD4^＋Foxp3^＋CD25^high，CD4^＋Foxp3^＋CD25^low和CD4^＋FoxD3^＋CD25^-细胞中CD127^low/-百分率分别为（93．8±3．5）％，（93．7±2．3）％，（92．0±2．1）％，三者之间差异无统计学意义（P〉0．05）；在CD4^＋细胞亚群中，当CD127^low／-时，Foxp3^＋在CD25^high，CD25^low和CD25^-中表达率分别为（91．4±2．6）％，（71．9±3．3）％，（9．0±2．2）％，三者之间差异均有统计学意义（P〈0．01）；SLE患者外周血CD4^＋CCR4^＋CD25^high T淋巴细胞百分率与SLEDAI呈显著负相关（r=-0．695，P〈0．001），狼疮肾炎患者（1．10±0．17）％显著低于SLE无肾炎组[（1．61±g0．23）％，P〈0．01]和健康对照组[（1．75±0．10）％，P〈0．01]；狼疮肾炎患者外周血CD4^＋CCR4^＋CD25^low/-T淋巴细胞百分率显著高于健康对照组[（11．5±2．3）％与（8．0±1．0）％，P〈0．01）]。结论初发SLE中异常升高的CD4^＋CD25^-Foxp3^＋T淋巴细胞的表型类似早期活化效应T淋巴细胞。可以用CD4＋CD25^highCD127^low/-T淋巴细胞替选CD4^＋CD25^highFoxp3^＋调节性T淋巴细胞。CCR4^＋调节性T淋巴细胞可能参与狼疮肾炎发病。

Objective To compare the phenotypes of abnormal CD4^＋CD25^-Foxp3^＋ T cells with traditional regulatory T cells （CD4^＋CD25^＋Foxp3^＋） in patients with untreated new-onset lupus （UNoL） and investigate their clinical relevance. Methods The expressions of surface markers （CD25, CD127, CCR4, GITR, CTLA-4） and intracellular marker（Foxp3） on the peripheral blood mononuclear cells from twenty-two UNoL patients were analyzed by flow cytometry analysis, and their clinical relevance were assessed. Results There were no significant differences between CD4^＋CD25^-Foxp3^＋ and CD4^＋CD25 ^＋Foxp3^- T ceils in the expressions of GITR, CTLA-4 and CCR4 （P〉0.05）, but they were significantly lower than those of CD4^＋CD25^＋ Foxp3^＋ T cells in UNoL patients （P〈0.01）. The percentages of CD127^low/- in CD4^＋Foxp3^＋CD25^high,CD4^＋Foxp3^＋ CD25^low and CD4^＋Foxp3^＋CD25^- T cells were （93.8±3.5）%, （93.7±2.3）% and （92.0±2.1）% respectively （P〉 0.05）, whereas the expressions of Foxp3 on CD4^＋CD127^low/- T subpoputations showed significant differences in CD4^＋CD127^low/-CD25^high （91.4±2.6）%, CD4^＋CD127^low/-CD25^low （71.9±3.3）% and CD4^＋CD127^low/-CD25^- （9.0± 2.2）% T cells（P〈0.01 ）. The frequeney of CD4^＋CCR4^＋CD25^high T cells correlated negatively with SLEDAI （r=-0.695, P〈0.001 ）, and it was significantly lower in lupus nephritis patients （1.10±0.17）% compared with SLE patients without nephritis [（1.61 ±0.23）%, P〈0.01] and healthy controls [（1.75±0.10）%, P〈0.01], furthermore, the frequency of CD4^＋CCR4^＋CD25^low/-T cells in lupus nephritis was significantly higher than that in healthy controls [ （ 11.5±2.3）% vs （8.0± 1.0）%, P〈0.01 ]. Conclusion The increased CD4^＋CD25^-Foxp3^＋ T cells in the Untreated Newonset Lupus （UNoL） patients mimic activated T effector cells. CD4^＋CD25^high-CD 127^low/- T cells can be used to isolate live CD4^＋CD25^highFoxp3^＋ regulatory T cells. CCR4^＋ regulatory T cells may be involved in the pathogenesis of lupus nephritis.