叶氏狼疮方诱导狼疮肾炎患者T细胞免疫耐受分子机制研究

Molecular Mechanism of Ye's Lupus Formula on Induction of Peripheral Blood T Lymphocytes Tolerance in the Patients with Lupus Nephritis

目的:探讨叶氏狼疮方在诱导狼疮肾炎(lupus nephritis,LN)患者外周血T淋巴细胞免疫耐受中有无作用。方法:密度梯度离心法提取LN患者及健康对照外周血单个核细胞(PBMCs),设立4个培养亚组:空白(blank)、植物血凝素(PHA)、狼疮方血清(lupus formula serum,LFS)、PHA+LFS组。双色标记流式细胞术检测PBMCs及其条件培养后CD3+T细胞表面分子CD28,CD40L,LFA-1a和CTLA-4的表达水平。多组间比较采用单因素方差分析。结果:活动性LN组外周血T细胞表面CD28、CD40L、LFA-1a和CTLA-4表达水平较健康对照组明显上调(P均<0.01);PHA可诱导健康对照T细胞表面CD28和CD40L表达明显上调(P均<0.01),但对其CTLA-4和LFA-1a的表达无明显影响(P均>0.05);而PHA对活动性LN组外周血T细胞表面上述指标表达均有明显上调作用(P均<0.05);叶氏狼疮方对健康人外周血T细胞表面上述指标的表达无明显影响,但可逆转活动性LN患者外周血T细胞表面的CD40L、LFA-1a过度表达(P均<0.05),对其CD28和CTLA-4过度表达无明显影响(P均>0.05);且叶氏狼疮方对PHA所诱导的活动性LN患者外周血T细胞表面CD28、CD40L和LFA-1a表达增加亦有明显抑制。结论:叶氏狼疮方可能主要通过下调T细胞CD40L和LFA-1a表达水平来诱导LN患者外周T细胞免疫耐受重建。

Objective：To investigate the effect of Ye＇s Lupus Formula on expression mode of costimulatory molecules group of T lymphocytes in the patients with lupus nephritis （LN）.Methods：The peripheral blood mononuclear cells of female LN patients and healthy females were abstracted by density gradient centrifugation.The Levels of CD28,CD40L,CTLA-4 and LFA-1a expression on the peripheral blood T lymphocytes were detected by double-colored flow cytometry.The difference of their means were tested by analysis of variance （ANOVA）.Results：Compared with the healthy controls,there were significantly higher level of CD28,CD40L,LFA-1a and CTLA-4 expression on the peripheral blood T cells in active LN patients（P〈0.01）.To the healthy controls,higher levels of CD28 and CD40L expression on the peripheral blood T cells could be induced by PHA（P〈0.01）,but not CTLA-4 and LFA-1a expression （P〈0.05）.To the LN patients,higher levels of all the above molecules expressions on the peripheral blood T cells could be induced by PHA（P〈0.05）.LFS could significantly inhibit over expression of LFA-1a and CD40L but for CD28 and CTLA-4 on the T cells of active LN patients（P〈0.05）,but did not affect the levels of all the above molecules expressions on the T cells of healthy controls.Further more,LFS could markedly inhibit all the above molecules＇ over expression induced by PHA on the T cells of active LN patients.Conclusion：Ye＇s Lupus Formula may be helpful to reestablish the immune tolerance of the peripheral blood T cells by down-regulation of CD40L and LFA-1a expression level on the peripheral blood T cells in LN patients.