中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2010年
7期
678-682
,共5页
王国兵%李成荣%杨军%温鹏强%贾实磊
王國兵%李成榮%楊軍%溫鵬彊%賈實磊
왕국병%리성영%양군%온붕강%가실뢰
川崎病%Foxp3%DNA甲基化%CD4+CD25+
川崎病%Foxp3%DNA甲基化%CD4+CD25+
천기병%Foxp3%DNA갑기화%CD4+CD25+
Kawasaki disease%Foxp3%DNA methylation%CD4+CD25+
目的 探讨急性期川崎病(KD)Foxp3基因甲基化状态及其在KD免疫发病机制中的作用.方法 急性期KD患儿30例,正常同年龄对照组18例,KD患儿分别于静脉丙种球蛋白(IVIG)治疗前后直接取血备检.甲基化特异性定量PCR(MSQP)检测CD4+T细胞Foxp3基因启动子、调节性T细胞(Tr)特异性甲基化区(TSDR)CpG岛甲基化状态;流式细胞术检测外周血CD4+CD25+Foxp3+Tr的比例;荧光定量PCR检测CD4+T细胞Foxp3、CTLA4、GITR、LAG3、CCR8、UBD、LGALS3等基因mRNA表达.结果 (1)急性期KD患儿CD4+T细胞Foxp3基因启动子CpG岛非甲基化水平明显低于同年龄对照组(P＜0.01),经IVIG治疗后明显升高(P＜0.01);TSDR区CpG岛非甲基化水平各组间差异无统计学意义(P＞0.05);(2)急性期KD患儿CD4+CD25+Foxp3+Tr细胞比例及CD4+T细胞Foxp3 mRNA表达水平明显低于正常对照组(P＜0.01),且与Foxp3基因启动子CpG岛非甲基化水平呈正相关(CD4+CD25+Foxp3+Tr:r=0.76,P＜0.01;Foxp3:r=0.89,P＜0.01),IVIG治疗后显著上升(P＜0.01);(3)急性期KD患儿CD4+CD25+Foxp3+Tr细胞相关因子CTLA4、GITR、LAG3和CCR8等基因mRNA水平显著降低(P＜0.01),IVIG治疗后均有不同程度的恢复(P＜0.01);Foxp3依赖性分子UBD和LGALS3表达水平亦低于正常对照组(P＜0.01),IVIG治疗后基本恢复至正常水平(P＜0.01).结论 急性期KD患儿Foxp3基因启动子非甲基化水平低下可能与KD免疫调节功能紊乱有关.
目的 探討急性期川崎病(KD)Foxp3基因甲基化狀態及其在KD免疫髮病機製中的作用.方法 急性期KD患兒30例,正常同年齡對照組18例,KD患兒分彆于靜脈丙種毬蛋白(IVIG)治療前後直接取血備檢.甲基化特異性定量PCR(MSQP)檢測CD4+T細胞Foxp3基因啟動子、調節性T細胞(Tr)特異性甲基化區(TSDR)CpG島甲基化狀態;流式細胞術檢測外週血CD4+CD25+Foxp3+Tr的比例;熒光定量PCR檢測CD4+T細胞Foxp3、CTLA4、GITR、LAG3、CCR8、UBD、LGALS3等基因mRNA錶達.結果 (1)急性期KD患兒CD4+T細胞Foxp3基因啟動子CpG島非甲基化水平明顯低于同年齡對照組(P＜0.01),經IVIG治療後明顯升高(P＜0.01);TSDR區CpG島非甲基化水平各組間差異無統計學意義(P＞0.05);(2)急性期KD患兒CD4+CD25+Foxp3+Tr細胞比例及CD4+T細胞Foxp3 mRNA錶達水平明顯低于正常對照組(P＜0.01),且與Foxp3基因啟動子CpG島非甲基化水平呈正相關(CD4+CD25+Foxp3+Tr:r=0.76,P＜0.01;Foxp3:r=0.89,P＜0.01),IVIG治療後顯著上升(P＜0.01);(3)急性期KD患兒CD4+CD25+Foxp3+Tr細胞相關因子CTLA4、GITR、LAG3和CCR8等基因mRNA水平顯著降低(P＜0.01),IVIG治療後均有不同程度的恢複(P＜0.01);Foxp3依賴性分子UBD和LGALS3錶達水平亦低于正常對照組(P＜0.01),IVIG治療後基本恢複至正常水平(P＜0.01).結論 急性期KD患兒Foxp3基因啟動子非甲基化水平低下可能與KD免疫調節功能紊亂有關.
목적 탐토급성기천기병(KD)Foxp3기인갑기화상태급기재KD면역발병궤제중적작용.방법 급성기KD환인30례,정상동년령대조조18례,KD환인분별우정맥병충구단백(IVIG)치료전후직접취혈비검.갑기화특이성정량PCR(MSQP)검측CD4+T세포Foxp3기인계동자、조절성T세포(Tr)특이성갑기화구(TSDR)CpG도갑기화상태;류식세포술검측외주혈CD4+CD25+Foxp3+Tr적비례;형광정량PCR검측CD4+T세포Foxp3、CTLA4、GITR、LAG3、CCR8、UBD、LGALS3등기인mRNA표체.결과 (1)급성기KD환인CD4+T세포Foxp3기인계동자CpG도비갑기화수평명현저우동년령대조조(P＜0.01),경IVIG치료후명현승고(P＜0.01);TSDR구CpG도비갑기화수평각조간차이무통계학의의(P＞0.05);(2)급성기KD환인CD4+CD25+Foxp3+Tr세포비례급CD4+T세포Foxp3 mRNA표체수평명현저우정상대조조(P＜0.01),차여Foxp3기인계동자CpG도비갑기화수평정정상관(CD4+CD25+Foxp3+Tr:r=0.76,P＜0.01;Foxp3:r=0.89,P＜0.01),IVIG치료후현저상승(P＜0.01);(3)급성기KD환인CD4+CD25+Foxp3+Tr세포상관인자CTLA4、GITR、LAG3화CCR8등기인mRNA수평현저강저(P＜0.01),IVIG치료후균유불동정도적회복(P＜0.01);Foxp3의뢰성분자UBD화LGALS3표체수평역저우정상대조조(P＜0.01),IVIG치료후기본회복지정상수평(P＜0.01).결론 급성기KD환인Foxp3기인계동자비갑기화수평저하가능여KD면역조절공능문란유관.
Objective To investigate the methylation status of Foxp3 gene and its roles in immunological pathogenesis of Kawasaki disease(KD). Methods Thirty children with KD and eighteen agematched healthy children consented to participate in this study. Quantitative methylation specific polymerase chain reaction(MSQP) was used to assess the methylation status of Foxp3 promoter and regulatory T cells specific demethylated region(TSDR) in CD4+ T cells. The proportion of CD4+ CD25 + Foxp3 + regulatory T (Tr) cells was analyzed by flow cytometry. Transcriptional levels of CD4+ CD25 + Foxp3 + Tr associating genes (Foxp3, CTLA4, GITR, LAG3 and CCR8 ) and Foxp3-dependent molecules (UBD and LGAIS3)were measured by real-time PCR. Results ( 1 ) Demethylation level of Foxp3 promoter in CD4 + T cells from patients with KD was lower significantly than that of health subjects( P ＜ 0.01 ), and increased significantly after treated with intravenous gamma globulin therapy(IVIG) (P ＜ 0.01 ). No difference of demethylation level of TSDR region was observed among all groups ( P ＞ 0. 05 ). ( 2 ) The proportion of CD4 + CD25 + Foxp3 + Tr in peripheral blood from patients with KD , as well as mRNA levels of Foxp3 gene in CD4+ T cells, was significantly lower than those of health subjects ( P ＜0. 01 ), and increases significantly after IVIG therapy (P ＜0.01 ). Significant positive correlations between demethylation level of Foxp3 promoter and the proportion of CD4 + CD25 + Foxp3 + Tr , or expression levels of Foxp3 in CD4 + T cells, were observed during acute phase of KD (CD4+CD25+ Foxp3+ Tr: r=0.76, P＜0. 01; Foxp3: r=0.89, P＜0. 01). (3)Transcription level of CD4+ CD25+ Foxp3+ Tr associating factors, such as CTLA4, GITR, LAG3 and CCR8, was significantly down-regulated in acute phase of KD(P＜0. 01 ), and up-regulated to some extent after treated with IVIG(P ＜0. 01 ). Expression levels of Foxp3-dependent molecules UBD and LGALS3 in CD4 + T cells decreased significantly during acute phase of KD (P ＜ 0.01 ), and basically recovered to the levels of health subjects( P ＜ 0.01 ). Conclusion Decrease of demethylation level of Fopx3 promoter is correlated with immune dysfunction in Kawasaki disease.
