异基因造血干细胞移植患者FOXP3 mRNA表达与移植物抗宿主病发生的关系

Correlation of FOXP3 mRNA expression to graft-versus-host disease occurrence in patients undergoing allogeneic hematopoietic stem cell transplantation

背景:转录因子FOXP3是调节性T细胞的特异性标志,对于造血干细胞移植后移植物抗宿主病的发生与FOXP3表达水平的关系目前仍存在争议。目的:观察异基因造血干细胞移植后患者外周血FOXP3mRNA的表达,初步探讨利用FOXP3mRNA表达水平的变化进行临床早期诊断移植物抗宿主病的可能性。设计:回顾性病例分析。对象:选取2001-03/2007-06在广东省人民医院血液科行异基因造血干细胞移植的白血病患者31例,男18例,女13例,平均年龄32岁。方法:所有患者均采用改良BU/CY联合FLU方案进行预处理,经深静脉输入单个核细胞数5.48(3.7～8.4)×108/kg,CD34+细胞数10.45(3.6～28.0)×106/kg。移植后出现移植物抗宿主病的患者在治疗前、治疗好转或缓解时留取外周血标本,尚无移植物抗宿主病的患者复查时采集外周血标本。主要观察指标:采用实时定量PCR技术监测外周血样本FOXP3mRNA表达及其与患者移植物抗宿主病的发生情况。结果:31例患者行异基因造血干细胞移植后,9例未出现移植物抗宿主病,9例发生急性移植物抗宿主病,13例出现慢性移植物抗宿主病。FOXP3表达水平的变化与急性移植物抗宿主病的临床演变相关,急性移植物抗宿主病发生时FOXP3水平较临床无移植物抗宿主病时明显降低(P=0.009),好转后FOXP3水平又逐步升高或恢复到基础水平;慢性移植物抗宿主病患者FOXP3的表达水平与无移植物抗宿主病患者相比无明显差异(P=0.107)。结论:急性移植物抗宿主病的发生与FOXP3表达降低有关,动态检测FOXP3mRNA表达水平的变化有可能为临床早期诊断急性移植物抗宿主病或预测其发生提供参考。实验数据未发现慢性移植物抗宿主病的发生与FOXP3表达有关。

BACKGROUND： Transcription factor FOXP3 is a specific marker of regulatory T cells. The correlation between graft-versus-host disease occurrence and FOXP3 expression is under debate following hematopoietic stem cell transplantation. OBJECTIVE： To observe peripheral blood FOXP3 mRNA expression following allogeneic hematopoietic stem cell transplantation and to primarily explore the possibility of clinical early diagnosis of graft-versus-host disease using FOXP3 mRNA expression. DESIGN： A retrospective case analysis. PARTICIPANTS： A total of 31 leukemia patients, who underwent allogeneic hematopoietic stem cell transplantation at the Department of Hematology, Guangdong Provincial People＇s Hospital from March 2001 to June 2007, were enrolled, including 18 males and 13 females, averagely 32 years old. METHODS： All patients underwent pretreatment of modified BU/CY and FLU scheme, and then infused with mononuclear cells at 5.48（3.7-8.4）×10^8/kg, and CD34＋ cells at 10.45（3.6-28.0）×10^5/kg via the deep vein. Peripheral blood samples were collected from posttransplantation patients with graft-versus-host disease before treatment, during improvement or remittance. Peripheral blood samples were harvested from patients without graft-versus-host disease during reexamination. MAIN OUTCOME MEASURES： Peripheral blood sample FOXP3 mRNA expression was quantified using real time quantitative poiymerase chain reaction, and the graft-versus-host disease occurrence was measured. RESULTS： Of the 31 patients undergoing allogeneic hematopoietic stem cell transplantation, 9 cases did not develop graft-versus-host disease, 9 cases developed acute graft-versus-host disease, and 13 cases developed chronic graft-versus-host disease. FOXP3 expression was correlated with clinical variation of acute graft-versus-host disease. FOXP3 expression was significantly decreased when acute graft-versus-host disease occurred compared with patients without graft-versus-host disease （P=0.009）, and gradually increased or reached to the basic levels following recovery. No significant differences in FOXP3 expression were detected between chronic graft-versus-host disease patients and no graft-versus-host disease patients （P=0.107）. CONCLUSION： The appearance of acute graft-versus-host disease is correlated with the decrease in FOXP3 expression. Dynamic examination of FOXP3 mRNA expression can provide a reference for diagnosing acute graft-versus-host disease early or predicting for the risk of acute graft-versus-host disease. Association of FOXP3 expression with chronic graft-versus-host disease was not observed in this study.