CD4^＋CD25^＋调节性T淋巴细胞与辅助性T淋巴细胞17在川崎病发病机制中的作用

Roles of CD4^＋ CD25^＋ regulatory T cells and T helper 17 cells in the pathogenesis of Kawasaki disease

目的观察川崎病（KD）患儿治疗前后外周血CD4^＋CD25^＋调节性T淋巴细胞（Treg）和辅助性T淋巴细胞17 （Th17）比例的变化，分析Treg/Th17细胞比例失衡在KD发病机制中的作用。方法收集2014年4至12月江西省儿童医院急性期KD患儿52例（KD组），同龄健康儿童34例（健康对照组）。KD患儿分别于丙种球蛋白（IVIG）、阿司匹林治疗前及治疗后热退3 d取清晨空腹静脉血，采用流式细胞术检测外周血CD4^＋CD25^＋ Treg和Th17细胞比例；酶联免疫吸附法检测血浆白细胞介素（IL）－6、IL－10、IL－17、IL－23及转化生长因子β（TGF－β）水平。结果急性期KD患儿外周血Treg细胞比例较健康对照组明显降低[（1.48±0.21）%比（5.13±0.32）%]，差异有统计学意义（t＝28.41，P〈0.05）；治疗后与治疗前相比显著升高[（4.71±0.36）%比（1.48±0.21）%]，差异有统计学意义（t＝－23.32，P〈0.05）。急性期KD患儿Th17细胞比例较健康对照组明显升高[（8.06±0.48）%比（2.65±0.50）%]，差异有统计学意义（t＝－23.47，P〈0.05）；治疗后与治疗前相比显著降低[（3.04±0.35）%比（8.06±0.48）%]，差异有统计学意义（t＝25.55，P〈0.05）。与健康对照组相比，急性期KD组患儿治疗前血清IL－6、IL－17和IL－23水平均显著升高[（34.53±0.53） ng/L比（10.88±0.83） ng/L、（57.05±0.78） ng/L比（14.29±0.98） ng/L、（45.18±1.52） ng/L比（18.25±1.08） ng/L]，差异均有统计学意义（t＝－72.36、－55.29、－43.27，均P〈0.05），IL－10及TGF－β水平显著降低[（14.29±0.64） ng/L比（29.57±0.87） ng/L、（16.88±0.90） ng/L比（38.83±0.84） ng/L] ，差异均有统计学意义（t＝42.24、53.51，均P〈0.05）；治疗后血清IL－6、IL－17和IL－23水平较治疗前均显著降低[（14.94±1.06） ng/L比（34.53±0.53） ng/L、（27.64±0.91） ng/L比（57.05±0.78） ng/L、（24.50±1.13） ng/L比（45.18±1.52） ng/L] ，差异均有统计学意义（t＝49.63、26.49、32.17，均P〈0.05），治疗后血清IL－10及TGF－β水平较治疗前显著升高[（23.01±0.61） ng/L比（14.29±0.64） ng/L、（33.47±0.82） ng/L比（16.88±0.90） ng/L]，差异均有统计学意义（t＝－29.54、－40.68，均P〈0.05）。结论Treg细胞与Th17细胞比例失衡可能是导致KD患儿免疫紊乱的重原因，体内相关细胞因子的变化参与该病的发病过程。

ObjectiveTo observe the changes in CD4^＋ CD25^＋ regulatory T cells（Treg） and T helper 17 cells（Th17） cells′ proportions in the peripheral blood in children with Kawasaki disease（KD） before and after the treatment, and to analyze the role of Treg/Th17 cell imbalance in the pathogenesis of KD.MethodsFifty-two children with acute KD（KD group） and 34 age-matched healthy children（healthy control group） were selected at Jiangxi Provincial Children′s Hospital from April to December of 2014.Morning peripheral vein blood was collected from 2 groups： one before the treatment by Immunoglobulin and Aspirin, and the other 3 days after defervescence treatment.Flow cytometry was used to detect proportions of Treg cells and Th17 cells in the peripheral blood.The enzyme linked immunosorbent assay was used to detect the levels of interleukin（IL）-6, IL-10, IL-17, IL-23 and transforming growth factor（TGF）-β.Results Proportion of Treg cells in the acute KD group was remarkably lower than that in the healthy control group [（1.48±0.21）%vs.（5.13±0.32）%, t=28.41, P〈0.05], but it was significantly increased after treatment, and there was a significant difference[（4.71±0.36）%vs.（1.48±0.21）%, t=-23.32, P〈0.05]. Proportion of Th17 cells in the acute KD group was markedly higher than that in the healthy control group[（8.06±0.48）%vs.（2.65±0.50）%, t=-23.47, P〈0.05], which was significantly decreased after treatment[（3.04±0.35）%vs.（8.06±0.48）%, t=25.55, P〈0.05]. Compared with the healthy control group, the levels of serum IL-6, IL-17, IL-23 in the acute KD group were significantly increased before treatment, and there were significant differences [（34.53±0.53） ng/L vs.（10.88±0.83） ng/L, t=-72.36; （57.05±0.78） ng/L vs.（14.29±0.98） ng/L, t=-55.29; （45.18±1.52） ng/L vs.（18.25±1.08） ng/L, t=-43.27; all P〈0.05], but after treatment the levels were significantly decreased[（14.94±1.06） ng/L vs.（34.53±0.53） ng/L, t= 49.63; （27.64±0.91） ng/L vs.（57.05±0.78） ng/L, t= 26.49; （24.50±1.13） ng/L vs.（45.18±1.52） ng/L, t= 32.17; all P〈0.05]. The levels of serum IL-10, TGF-β in the acute KD group significantly decreased than those of the healthy control group, and there were significant differences [（14.29±0.64） ng/L vs.（29.57±0.87） ng/L, t=42.24; （16.88±0.90） ng/L vs.（38.83±0.84） ng/L, t=53.51; all P〈0.05 ], but after treatment the levels were significantly increased, and there were significant differences [（23.01±0.61） ng/L vs.（14.29±0.64） ng/L, t=-29.54; （33.47±0.82） ng/L vs.（16.88±0.90） ng/L, t=-40.68; all P〈0.05 ].Conclusion Imbalance between Treg cells and Th17 cells may be an important cause for the immune disorder of KD, the changes in related cytokines are involved in the pathogenesis of KD.