摘要
目的 探讨急性期川崎病患儿CD4^＋CD25^highFOXP3^＋调节性T细胞亚群改变及其在川崎病免疫发病机制中的作用.方法 急性期川崎病患儿36例,同年龄健康对照组32名,川崎病患儿治疗前、后取血备检.采用流式细胞术检测外周血CD4^＋CD25^highFOXP3^＋、可诱导性T细胞共刺激分子（ICOS）^＋调节性T细胞、ICOS-调节性T细胞比例及IL-10、TGF-β、IL-35p35、IL-35EBI3、TGF-βRⅡ、ICOS、CD28蛋白表达水平;实时荧光定量PCR检测CD4^＋T细胞Smad3/4、TGF-β诱导的早期反应基因1（TIEG1）、痒同源物E3泛素蛋白连接酶（hch）等mRNA表达;ELISA检测血浆中TGF-β浓度.采用t检验进行统计分析.结果 ①急性期川崎病患儿调节性T细胞比例[（3.2±1.8）％]明显低于对照组[（7.3±2.9）％,t=6.9,P＜0.05],其中ICOS^＋调节性T细胞和ICOS^-调节性T细胞比例[（2.3±1.0）％,（0.9±0.3）％]均明显低于健康对照组[（4.7±1.4）％,（2.6±0.9）％,t=11.7,9.8;P均＜0.05],且ICOS＋调节性T细胞/ICOS^-调节性T细胞比值（2.6±0.7）高于对照组[（1.8±0.5）,t=9.4,P＜ 0.05],治疗后均明显恢复[（5.9±2.3）％,（3.9±1.6）％,（2.0±0.8）％,（1.9±0.5）％;t=5.5,5.0,8.2,4.9;P均＜0.05];②急性期川崎病患儿ICOS＋调节性T细胞FOXP3、IL-10、TGF-β、IL-35p35和IL-35EBI3表达水平明显低于对照组（t=5.5,4.8,8.0,6.6,9.6;P均＜0.05）,而ICOS-调节性T细胞FOXP3、mTGF-β表达亦低于对照组（t=7.5,10.0;P均＜0.05）,治疗后上述分子表达显著上调（t=3.7,3.7,8.5,6.1,7.7,5.3,7.0;P均＜0.05）;③急性期川崎病患儿血浆TGF-β浓度及CD4＋T细胞TGF-βRⅡ、Smad3、Smad4、TIEG1、Itch表达水平明显低于对照组,调节性T细胞表面ICOS、CD28表达亦低于健康对照组,治疗后均明显恢复.结论 TGF-β、ICOS、CD28信号异常可能是导致急性期川崎病患儿CD4^＋CD25^highFOXP3^＋调节性T细胞及其亚群比例失调的重要因素.
Objective To investigate the changes and significances of CD4^＋CD25^highFOXP3^＋ regulatory T cells （Treg） and its subsets in children with acute Kawasaki disease （KD）.Methods Thirty-six children with KD and thirty-two age-matched healthy donors were studied.Flow cytometry was performed to evaluate the proportions of Treg and its subsets [inducible T-cell co-stimulator （ICOS）^＋ Treg and ICOS-Treg],and the expression levels of associated molecules such as interleukin （IL）-10,transform growth factor （TGF）-β,IL-35,TGF-βR Ⅱ,ICOS and CD28 were tested.Transcription levels of Smad family member （Smad） 3/4,TGF beta inducible early response gene 1 （TIEG1） and itchy E3 ubiquitin protein ligase （Itch） in CD4^＋ T cells were determined by quantitative real-time polymerase chain reaction （PCR）.Plasma concentrations of TGF-β were measured by enzyme-linked immunosorbent assay.Independent-samples t-test was used as the statistical method in this study.Results ① The proportions of Treg and its subsets （ICOS^＋ Treg and ICOS-Treg） in children with acute KD [（3.2±1.8）%,（2.3±1.0）%,（0.9±0.3）%] were lower than those in healthy controls [（7.3± 2.9）%,（4.7±1.4）%,（2.6±0.9）%; t=6.9,11.7,9.8; P〈0.05],which resulted in a higher ratio of ICOS^＋ Treg/ICOS-Treg [（2.6±0.7） vs （1.8±0.5）,t=9.4,P〈0.05],and restored remarkably after therapy [（5.9±2.3）%,（3.9±1.6）%,（2.0±0.8）%,（1.9±0.5）%; t=5.5,5.0,8.2,4.9; P〈0.05].② Expression levels of FOXP3,IL-10,TGF-β,IL-35p35 and IL-35EBI3 in ICOS＋ Treg of patients with acute KD were found to be lower than those in healthy controls （t=5.5,4.8,8.0,6.6,9.6; P〈0.05）.Meanwhile,FOXP3 and membrane bound TGF-β on ICOS-Treg in children with acute KD were lower than those of healthy subjects （t=7.5,10.0; P〈0.05）,although expression levels of all the molecules mentioned before were elevated after therapy （t=3.7,3.7,8.5,6.1,7.7,5.3,7.0; P〈0.05）.③ Plasma TGF-β concentration and e
出处
《中华风湿病学杂志》
CAS
CSCD
北大核心
2015年第2期91-96,共6页
Chinese Journal of Rheumatology
基金
国家自然科学基金，广东省医学科学技术研究基金，深圳市科学技术项目
