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目的 检测输血相关急性肺损伤(TRALI)患者外周血CD4+CD25highFoxp3+调节性T淋巴细胞(Treg)的计数及其功能基因叉头框蛋白3 (Foxp3) mRNA的表达水平,以及探讨其与疾病预后的关系.方法 收集2008年6月至2011年11月期间健康体检者(对照组,n=30)和重症监护病房收治的TRALI患者[TRALI组,n=26,TRALI组按预后又分为死亡组(n=5)和存活组(n=21)]外周抗凝血,采用流式细胞术分析各组T细胞亚群.采用四色流式细胞术以Foxp3-FITC/CD25-PE/CD4-PerCP/CD3-PC7抗体组合检测各组受试者外周血CD4+CD25highFoxp3 +Treg的计数,比较死亡组和存活组患者外周血CD4+CD25highFoxp3+Treg的水平.实时荧光定量PCR技术检测特异性转录因子Foxp3 mRNA的表达水平,并分析患者外周血CD4+CD25highFoxp3+Treg的比例与Foxp3mRNA的表达水平的关系.结果 TRALI患者与对照组健康人间CD3+T细胞、CD3+CD4+T细胞计数的差异无统计学意义,而CD3+CD8+T细胞则较对照组增加(P＜0.05),CD4+/CD8+比值降低(P＜0.05).死亡组CD3+CD8+T细胞比例高于存活组(P＜0.05),CD3+T细胞、CD3+CD4+T细胞比例和CD4+/CD8+比值则低于存活组(均P＜0.05).TRALI患者外周血CD4+CD25highFoxp3+Treg计数为(8.86±3.14)％,明显高于健康对照组(5.67±2.43)％(P＜0.05).TRALI死亡组患者外周血CD4+CD25highFoxp3+Treg计数为(11.23±3.79)％,显著高于存活组患者(7.69±3.21)％(P＜0.05).TRALI患者外周血Foxp3 mRNA的表达水平为(3.77±2.73)×105拷贝/μg,显著高于对照组(0.43±0.21) ×105拷贝/μg(P＜0.05).TRALI患者外周血CD4+CD25highFoxp3+ Treg计数与Foxp3 mRNA的表达水平呈正相关(r=0.513 1,P＜0.05).结论 TRALI患者外周血CD4+C D25highFoxp3+ Treg和Foxp3mRNA的变化可能是导致TRALI疾病发展的关键因素之一,与疾病预后有密切关系.

Objective To investigate the count of peripheral blood CD4+CD25highFoxp3+ regulatory T cells and mRNA expression of its functional gene,fork-head box protein 3 (Foxp3),in patients with transfusion-related acute lung injury (TRALI),and to explore their associations to disease prognosis.Methods Anticoagulated peripheral blood samples were collected from health checkup subjects (control group,n=30) and TRALI patients in ICU [TRALI group,n=26,comprising a death sub-group (n=5) and a survival sub-group (n=21)] of our hospital between June 2008 and November 2011.Routine flow cytometry was used to analyze the T lymphocyte subsets in the blood.Four-color flow cytometry with Foxp3-FITC/CD25-PE/CD4-PerCP/CD3-PC7 as the combination of antibodies was further used to determine the count of peripheral blood CD4 + CD2highFoxp3 + Treg.The level of peripheral CD4 + CD25highFoxp3 + Treg was compared between the death and survival sub-groups of TRALI patients.Level of Foxp3 mRNA expression was measured with a real-time quantitative PCR.The correlation between the percentage of peripheral CD4 + CD25highFoxp3+Treg and the Foxp3 mRNA expression was analyzed.Results Between TRALI patients and healthy controls,there were no statistical differences in peripheral cell counts of CD3+ and CD3+CD4+ T cells,but there were higher level of CD3+CD8+ T cells and lower ratio of CD4+/CD8+ (both P＜0.05) in TRALI patients.The death sub-group showed higher percentage of CD3+CD8+ T cells (P＜0.05),lower percentage of CD3+ and CD3+CD4+ T cells,and lower ratio of CD4+/CD8+ compared with the survival sub-group (all P＜0.05).TRALI patients had statistically higher level of C D4 + CD25highFoxp3 + Treg [(8.86 ± 3.14)％] than healthy controls [(5.67±2.43) ％] (P＜0.05),and patients in the dead sub-group had statistically higher level of CD4+ CD25highFoxp3+Treg [(11.23±3.79)％] than those in the survival sub-group [(7.69±3.21)％] (P＜0.05).In addition,there was a positive correlation between the level of peripheral CD4+CD25high Foxp3+ Treg and Foxp3 mRNA expression (r=0.513 1,P＜0.05) in TRALI patients.Conclusion The changes in the count of peripheral blood CD4+CD25highFoxp3+ Treg and Foxp3 mRNA may be a critical factor leading to TRALI in the patients,which is closely associated with the disease prognosis.