CAJ | 학술논문

目的探讨sIL-2R血浓度对急性期川崎病（ KD）患儿调节性T细胞（ Treg）的影响。方法急性期KD患儿33例，正常同龄对照儿童14例。流式细胞术检测外周血CD4＋CD25＋Foxp3＋Treg细胞的比例和CD4＋CD25＋T细胞磷酸化STAT5（pSTAT5）蛋白平均荧光强度（MFI）；流式微球阵列术（CBA）检测血浆sIL-2R、IL-2、IL-7、IL-15的浓度；荧光定量PCR（real-time PCR）检测CD4＋CD25＋T细胞Foxp3、GITR、CTLA-4、IL-2Rα、IL-2Rβ、IL-2Rγ和CD 4＋CD25-T 细胞 IL-17A、RoR-γt 等基因mRNA表达。结果（1）急性期KD患儿CD4＋CD25＋Foxp3＋Treg细胞比例及相关分子Foxp3、GITR、CTLA-4 mRNA表达明显低于同龄对照组（P＜0．05），Th17细胞相关因子IL-17A、ROR-γt mRNA表达明显增高（P＜0．05），IVIG治疗后呈不同程度的恢复（P＜0．05）。（2）急性期KD患儿CD4＋CD25＋T细胞pSTAT5蛋白水平显著下调（P＜0．05），IVIG治疗后明显上调（P＜0．05）。（3）急性期KD患儿血浆sIL-2R浓度显著增高（P＜0．05），IVIG治疗后下降（P＜0．05）；其中KD合并冠脉损伤组（KD-CAL＋）明显高于无冠脉损伤组（KD-CAL-）（P＜0．05）；IL-2、IL-7、IL1-5浓度无明显改变（P＞0．05）。（4）急性期KD患儿CD4＋CD25＋T细胞IL-2Rα、IL-2Rβ基因mRNA表达明显低于同龄对照组（P＜0．05），IVIG治疗后呈不同程度的升高（ P＜0．05）；IL-2Rγ基因mRNA表达无明显改变；sIL-2R血浓度与IL-2RβmRNA、pSTAT5及Foxp3 mRNA表达呈负相关（ P＜0．05）；pSTAT5与Foxp3 mRNA表达呈正相关（ P＜0．05）。结论血浆sIL-2R明显增高可致IL-2/STAT5信号途径传导异常，这可能是导致急性期KD Treg细胞下调的因素之一。
목적탐토sIL-2R혈농도대급성기천기병（ KD）환인조절성T세포（ Treg）적영향。방법급성기KD환인33례，정상동령대조인동14례。류식세포술검측외주혈CD4＋CD25＋Foxp3＋Treg세포적비례화CD4＋CD25＋T세포린산화STAT5（pSTAT5）단백평균형광강도（MFI）；류식미구진렬술（CBA）검측혈장sIL-2R、IL-2、IL-7、IL-15적농도；형광정량PCR（real-time PCR）검측CD4＋CD25＋T세포Foxp3、GITR、CTLA-4、IL-2Rα、IL-2Rβ、IL-2Rγ화CD 4＋CD25-T 세포 IL-17A、RoR-γt 등기인mRNA표체。결과（1）급성기KD환인CD4＋CD25＋Foxp3＋Treg세포비례급상관분자Foxp3、GITR、CTLA-4 mRNA표체명현저우동령대조조（P＜0．05），Th17세포상관인자IL-17A、ROR-γt mRNA표체명현증고（P＜0．05），IVIG치료후정불동정도적회복（P＜0．05）。（2）급성기KD환인CD4＋CD25＋T세포pSTAT5단백수평현저하조（P＜0．05），IVIG치료후명현상조（P＜0．05）。（3）급성기KD환인혈장sIL-2R농도현저증고（P＜0．05），IVIG치료후하강（P＜0．05）；기중KD합병관맥손상조（KD-CAL＋）명현고우무관맥손상조（KD-CAL-）（P＜0．05）；IL-2、IL-7、IL1-5농도무명현개변（P＞0．05）。（4）급성기KD환인CD4＋CD25＋T세포IL-2Rα、IL-2Rβ기인mRNA표체명현저우동령대조조（P＜0．05），IVIG치료후정불동정도적승고（ P＜0．05）；IL-2Rγ기인mRNA표체무명현개변；sIL-2R혈농도여IL-2RβmRNA、pSTAT5급Foxp3 mRNA표체정부상관（ P＜0．05）；pSTAT5여Foxp3 mRNA표체정정상관（ P＜0．05）。결론혈장sIL-2R명현증고가치IL-2/STAT5신호도경전도이상，저가능시도치급성기KD Treg세포하조적인소지일。
Objective To investigate the effects of sIL-2R in plasma on regulatory T cells (Treg) in children with Kawasaki disease ( KD ) .Methods Thirty-three children with KD and fourteen age-matched healthy children were enrolled in this study .The proportions of CD4+CD25+Foxp3+Treg cells in pe-ripheral blood and mean fluorescence intensity (MFI) of phosphorylated-STAT5 (pSTAT5) protein in CD4+CD25+T cells were analyzed by flow cytometry .The concentrations of sIL-2R, IL-2, IL-7 and IL-15 in plas-ma were measured by cytometric bead array ( CBA ) .Real-time PCR was performed to detect the gene ex-pressions of Foxp3, GITR, CTLA4, IL-2Rα, IL-2Rβand IL-2Rγat mRNA level as well as the expressions of IL-17A and ROR-γt at mRNA level in CD4+CD25-T cells.Results (1) Compared with the control group, the proportions of CD4+CD25+Foxp3+Treg cells in peripheral blood from patients with KD and the expressions of associated factors including Foxp 3, GITR and CTLA-4 at mRNA level were significantly down-regulated (P<0.05).However,the expressions of IL-17A and ROR-γt at mRNA level in Th17 cells were markedly up-regulated (P<0.05), which could be recovered to some extent after treatment with IVIG (P<0.05).(2)The expressions of pSTAT5 protein in CD4+CD25+T cells from patients with acute KD were sig-nificantly decreased (P<00.5 ), but increased with IVIG intervention (P<0.05).(3)The concentrations of sIL-2R in plasma were elevated during acute KD (P<0.05), but decreased after treatment with IVIG (P<0.05).Moreover, KD patients with coronary artery lesion ( KD-CAL+) presented a high level of sIL-2R than those without coronary artery lesion (KD-CAL-) (P<0.05), but there was no significant difference in the concentrations of IL-2, IL-7 and IL-15 in plasma between two groups (P>0.05).(4)The expressions of IL-2Rαand IL-2Rβat mRNA level in CD4+CD25+T cells from patients with acute KD were lower than those of the healthy subjects (P<0.05), but up-regulated to some extent with IVIG treatment (P<0.05).There was no significant change in the expression of IL-2Rγat mRNA level (P>0.05).The concentrations of sIL-2R in plasma were negatively correlated with the expressions of IL-2Rβand Foxp3 at mRNA level and pSTAT5 at protein level (P<0.05).The expression of pSTAT5 protein had positive correlation with the ex-pression of Foxp3 at mRNA level (P<0.05).Conclusion Aberrant IL-2/STAT5 signaling pathway media-ted by significantly increased concentration of sIL-2R in plasma might be one of the factors leading to down-regulation of Treg cells in patients with acute KD .