免疫抑制剂对Graves病患者免疫调节细胞的体内外作用研究

The effects of immunosuppressors on immunoregulatory cells in patients with Graves disease in vitro and in vivo

目的观察Graves病(Graves disease,GD)患者外周血免疫调节细胞在体外对地塞米松(DEX)、硫唑嘌呤(AZA)及环磷酰胺(CTX)等免疫抑制剂的反应及其体内在甲状腺局部注射DEX后的变化,探讨DEX等免疫抑制剂在甲状腺局部注射治疗GD过程中对免疫调节系统的作用。方法 1)用DEX、AZA及CTX等免疫抑制剂处理GD患者外周血单个核细胞,采用流式细胞术检测调节性T细胞(Treg)、辅助性T细胞(Th)、调节性B细胞(Breg)、树突状细胞(DC)等免疫细胞亚群变化。2)21例抗甲状腺药物治疗维持期的GD患者,甲状腺局部注射DEX,治疗前后分别检测外周血Treg、Th、Breg、DC等免疫细胞亚群变化。结果 1)免疫抑制剂处理后,各组Treg水平均减少(P<0.05),DEX组Th2细胞减少(P<0.05),CTX组Th1细胞增加(P<0.05);DEX组Breg增多(P<0.05),其余2组无明显统计学差异;各组DC细胞中,mDC均明显升高伴pDC下降,DEX、CTX组(P<0.05),AZA组(P<0.01),DEX组成熟型CD83+及HLA-DR+DC均下降(P<0.05),其余2组无明显统计学差异。2)甲状腺局部注射DEX后,与注射前相比,GD患者外周血中Treg细胞增多(P<0.05),Th2细胞减少(P<0.05),Th1细胞无明显变化(P>0.05);Breg比例升高(P<0.05);DC亚群中,mDC升高伴pDC下降(P<0.05),成熟型CD83+及HLA-DR+DC比例均下降(P<0.05)。结论 DEX能有效调节GD患者外周血Treg、Breg及DC等调节性细胞及其亚群水平,重建Th1/Th2平衡,可作为甲状腺局部注射免疫抑制剂治疗GD的首选用药。

In order to explore the roles of DEX and other immunosuppressors in immunoregulatory system of Graves disease（GD）, we investigate the responses of immune regulatory system to dexamethasone（DEX）,azathioprine（AZA） and cyclophosphamide（CTX） in peripheral blood of GD patients in vitro, and the immune regulatory effects of intrathyroid injection of DEX in vivo. Peripheral blood mononuclear cells（PBMC） of GD patients were separated and treated with DEX, AZA and CTX respectively, and then flow cytometry was used to detect the changes of regulatory T（Treg） cells, helper T（Th） cells, regulatory B（Breg） cells, dendritic cells（DC） and their subsets. In addition, twenty-one patients with GD who maintained antithyroid drugs as a background treatment were treated by an intrathyroid injection of DEX, and the Treg, Th, Breg cells, DC and their subsets were detected.The results showed that the Treg cells decreased significantly after the treatment of DEX, AZA or CTX, compared with controls; Th2 cells decreased in DEX group, and Th1 cells increased in CTX group; the Breg cells increased after DEX treatment but there was no significant difference between AZA and CTX groups. In DC subsets, mDC increased but pDC decreased in all three groups. Both CD83＋and HLA-DR＋DC decreased in DEX group, but there was no significant change between AZA and CTX groups. Furthermore, after intrathyroid injection of DEX, Treg,Breg cells increased, Th2 cells decreased in patients with GD, but there was no significant difference in Th1 cells before and after treatment. As for DC subsets, DEX injection increased mDC, but decreased pDC, CD83＋DC and HLA-DR＋DC. The results suggested that DEX could regulate the numerical impairments of Treg, Breg, DC and their subsets, and rebuild the Th1/Th2 balance, thus DEX might be the first choice in intrathyroid injection treatment for GD.