CAJ | 학술논문

目的观察不同免疫状态下脓毒症婴幼儿外周血CD4+CD25+Foxp3high调节性T细胞(Treg细胞)及相关分子的变化,探讨婴幼儿脓毒症免疫功能紊乱的可能机制.方法分别收集2007年5月至2007年11月深圳市儿童医院重症监护室收治的婴幼儿脓毒症36例血液标本,另选16例健康同龄儿童作为正常对照进行前瞻性研究;排除既往患有自身免疫性疾病、免疫缺陷病、遗传代谢病及肿瘤的患儿,排除近6个月曾使用影响免疫功能的药物.本研究获得深圳市儿童医院伦理委员会的同意.以外周血CD14+单核细胞HLA-DR表达>30%或<30%为阈值,将患儿分为免疫激活组(DR-H组)和免疫抑制组(DR-L组),用流式细胞术检测CD14+单核细胞HLA-DR表达率,CD4+CD25+Foxp3highTreg细胞比例;实时荧光定量PCR(Real time-PCR)检测CD4+T细胞Foxp3、CTLA-4、GITR、IL-10mRNA表达.统计方法采用单因素方差分析,P<0.05为差异具有统计学意义.结果急性期DR-L组CD4+CD25+Foxp3highTreg细胞比例明显高于对照组及DR-H组(P<0.05).DR-L组Foxp3、CTLA-4、IL-10等相关分子基因表达高于对照组及DR-H组(P<0.05),DR-L组GITR基因表达高于DR-H组.结论 CD4+CD25+Foxp3highTreg细胞数量异常增加可能与婴幼儿脓毒症免疫抑制状态有关.
목적 관찰불동면역상태하농독증영유인외주혈CD4+CD25+Foxp3high조절성T세포(Treg세포)급상관분자적변화,탐토영유인농독증면역공능문란적가능궤제.방법 분별수집2007년5월지2007년11월심수시인동의원중증감호실수치적영유인농독증36례혈액표본,령선16례건강동령인동작위정상대조진행전첨성연구;배제기왕환유자신면역성질병、면역결함병、유전대사병급종류적환인,배제근6개월증사용영향면역공능적약물.본연구획득심수시인동의원윤리위원회적동의.이외주혈CD14+단핵세포HLA-DR표체>30%혹<30%위역치,장환인분위면역격활조(DR-H조)화면역억제조(DR-L조),용류식세포술검측CD14+단핵세포HLA-DR표체솔,CD4+CD25+Foxp3highTreg세포비례;실시형광정량PCR(Real time-PCR)검측CD4+T세포Foxp3、CTLA-4、GITR、IL-10mRNA표체.통계방법채용단인소방차분석,P<0.05위차이구유통계학의의.결과 급성기DR-L조CD4+CD25+Foxp3highTreg세포비례명현고우대조조급DR-H조(P<0.05).DR-L조Foxp3、CTLA-4、IL-10등상관분자기인표체고우대조조급DR-H조(P<0.05),DR-L조GITR기인표체고우DR-H조.결론 CD4+CD25+Foxp3highTreg세포수량이상증가가능여영유인농독증면역억제상태유관.
Objective To study the change of CD4+ CD25+ Foxp3high regulatory T cells (Treg cells) and the molecules associated with Treg cells in different immune status in infant with sepsis, and to further clarity the pathogenesis of disturbed immune function in infant with sepsis. Method Totally 36 sepsis infants admitted in In-tensive Care Unit of Shenzhen Children' s Hospital from May 2007 to November 2007 and 16 age-matched healthy infants were collected for prospective study, after excluding autoimmune disease, immunodeficiency, inherited metabolic disorders, tumor, and drug-treatment that could affect immune function during lately 6 months. The study was approved by Ethics Committee of Shenzhen Children's Hospital. The 36 infants with sepsis were divided into two groups according to expression levels of HLA-DR in CD14-positive cells: DR-H group was defined as patients with HLA-DR > 30%, while DR-L group was defined as patients with HLA-DR < 30%. Expression levels of HLA-DR in CD14-positive cells and the proportion of Treg cells were analyzed by flow cytometry. Real-time PCR were used to evaluate the mRNA levels of Foxp3, CTLA-4,GITR, and IL-10 in CD4-posidve ceils. Statistical analysis was performed by one-way Anova. There was statistical difference with P < 0.05. Results The proportion of Treg cells in DR-L group was found to be significantly higher than that in healthy control or DR-H group (P <0.05).Compared with healthy control group or DR-H group, transcriptional levels of Foxp3, CTLA-4 and IL-10were significantly increased in DR-L group (P <0.05). The levels of GITR mRNA in DR-L group were detected to be higher than those in DR-H group (P < 0.05). Conclusions Aberrant increased proportion of Treg cells may be associated with suppressed immune status in infant with sepsis.