风湿性疾病患者外周血CD4^(+)T细胞亚群特征及其对免疫调节联合治疗的反应

The status of peripheral CD4^(+)T subsets in patients with rheumatism and their changes after immuno-modulatory combination therapies

目的测定风湿性疾病患者外周血CD4^(+)T细胞亚群水平,探讨免疫调节联合方案治疗前后的变化。方法回顾性分析了6395例风湿性疾病患者和206名健康者外周血CD4^(+)T细胞亚群水平,包括RA、SLE、AS、PsA、SSc、pSS、白塞病、PM/DM、痛风和血管炎患者,其中一部分患者接受了低剂量IL-2、雷帕霉素、二甲双胍、维甲酸、辅酶Q10等免疫调节药物治疗。用流式细胞术检测受试者外周血辅助细胞(Th)1、Th2、Th17和调节性T细胞的绝对计数。采用独立样本t检验和配对样本t检验分别比较患者与健康对照组、治疗前后外周血CD4^(+)T细胞亚群水平,P<0.05为差异有统计学意义。结果与健康对照组相比,患者的调节性T细胞水平明显偏低[(31±15)个/μl与(27±17)个/μl,t=3.407,P<0.01;t=3.407,P<0.01],Th17/Treg显著升高[(0.3±0.2)与(0.4±0.7),t=-9.508,P<0.01]。其中,AS[(10±8)个/μl,t=-5.403,P<0.01]、PsA[(11±11)个/μl,t=-3.829,P<0.01]、SSc[(7±6)个/μl,t=3.114,P<0.01]、BD[(11±9)个/μl,t=-4.774,P<0.01]和痛风[(11±9)个/μl,t=-4.604,P<0.01]患者的Th17高于健康对照组(8±5)个/μl,RA[(28±15)个/μl,t=3.032,P<0.01]、SLE[(21±21)个/μl,t=6.836,P<0.01]、AS[(28±15)个/μl,t=2.216,P<0.05]、SSc[(27±16)个/μl,t=3.698,P<0.05]、BD[(27±17)个/μl,t=2.502,P<0.05]、DM/PM[(27±22)个/μl,t=2.974,P<0.01]和痛风[(28±15)个/μl,t=2.079,P<0.05]患者调节性T细胞均低于正常[(31±15)个/μl]。经过免疫调节联合治疗后,患者的CD4^(+)T细胞亚群明显改变,尤其以调节性T细胞的增加为著,因调节性T细胞的增长速率快于其他效应T细胞,致使相应细胞如Th17/调节性T细胞回降,其中SLE较为显著[(0.6±1.0)与(0.5±0.4),t=3.157,P<0.01]。结论风湿性疾病是一类以淋巴细胞亚群失衡为主要特征的自身免疫紊乱的炎症,其中以调节性T细胞相对不足最为显著。免疫调节药物(IMiDs)治疗的免疫疗法可相对特异地促进调节性T细胞增殖,诱导和恢复患者的自身免疫耐受,有望为风湿性疾病的治疗提供新思路。

Objective To examinethe absolute numbers of cluster of differentiation(CD4)^(+)T cell subsets in peripheral blood of patients with rheumatism and further to develop a new immunomodulatory therapies which aimed to restore their imbalanced CD4^(+)T lymphocyte subpopulation.Methods A total of 6395 rheumatic patients[4430 females,1965 males,mean age(49±15)years]and 206 healthy controls(HCs)were enrolled in this retrospective cross-sectional study,which included rheumatoid arthritis(RA),systemic lupus erythematosus(SLE),ankylosing spondylitis(AS),Psoriatic arthritis(PsA),systemic sclerosis(SSc),primary Sjögren′s syndrome(pSS),Beçhet's disease(BD),dermatomyositis/polymyositis(DM/PM),gout and vasculitis.Some patients received treatment combined with immunoregulatory drugs(IMiDs)such as low-dose interleukin(IL)-2,rapamycin,metformin,retinoic acid and coenzyme Q10.The absolute numbers of T helper cell(Th)1,Th2,Th17 and regulatory T cell(Treg)in peripheral blood(PB)of these individuals were measured by Flow Cytometery(FCM).Independent sample t test and paired sample t test were used to compare the levels of CD4^(+)T cell subsets in PB of patients and HCs,before and after treatment respectively,and P<0.05 was considered statistically significant.Results Compared with HCs,the mean absolute number of Treg was significantly decreased[(31±15)cell/μl vs(27±17)cell/μl,t=3.407,P<0.01]and the ratio of Th17/Treg was increased in all patients[(0.3±0.2)vs(0.4±0.7),t=-9.508,P<0.01].There was a significant increase in the number of Th17 in patients with AS[(10±8)cell/μl,t=-5.403,P<0.01],PsA[(11±11)cell/μl,t=-3.829,P<0.01],SSc[(7±6)cell/μl,t=3.114,P<0.01],BD[(11±9)cell/μl,t=-4.774,P<0.01]and gout[(11±9)cell/μl,t=-4.604,P<0.01],and we observed lower level of Treg in patients with RA[(28±15)cell/μl,t=3.032,P<0.01],SLE[(21±21)cell/μl,t=6.836,P<0.01],AS[(28±15)cell/μl,t=2.216,P<0.05],SSc[(27±16)cell/μl,t=3.698,P<0.05],BD[(27±17)cell/μl,t=2.502,P<0.05],DM/PM[(27±22)cell/μl,t=2.974,P<0.01]and gout[(28±15)cell/μl,t=2.079,P<0.05].After IMiDs combination treatment,the levels of CD4^(+)T subsets were increased.Interestingly,the expansion of Treg was much more dramatical than those of other effector T cells,resulting in a decrease in ratios of Th17/Treg,especially in SLE[(0.6±1.0)vs(0.5±0.4),t=3.157,P<0.01].Conclusion Impaired balance of pro-and anti-inflammatory immune cells,especially insufficiency of Treg,might be a cornerstone of the pathogenesis of rheumatism.The new immunomodulatory therapies could relatively specifically promote Treg proliferation and restored patients'autoimmune tolerance,which isexpected to provide a new strategy for the treatment of rheumatism.