异基因造血干细胞移植后aGVHD与调节性T细胞、IL-35的相关性  

Relationship between regulatory T cells,IL-35 and aGVHD after allo-HSCT

作　　者：孙元欣[1] 秦雪梅[1] 刘希民[2] 王立志[2] 刘传方[1] 刘璐[1] 

机构地区：[1]山东大学齐鲁医院血液科,济南250012 [2]济南军区总医院血液科,济南250031

出　　处：《山东大学学报（医学版）》2013年第3期89-94,共6页Journal of Shandong University:Health Sciences

基　　金：山东大学自主创新基金(2009TS065);济南市留学回国人员创业计划(20100407)

摘　　要：目的探索异基因造血干细胞移植(allo-HSCT)后发生急性移植物抗宿主病(aGVHD)的患者外周血中调节性T细胞及其相关转录因子、细胞因子的水平,以期获得能够有效预测患者发生aGVHD的生物学指标。方法外周血样品采自接受造血干细胞移植术的患者和健康人,其中aGVHD阳性组13例,aGVHD阴性组10例,正常对照组21例。采用流式细胞术检测各组外周血中CD4+CD25+Foxp3+Treg细胞占CD4+T淋巴细胞的百分比,应用实时定量PCR检测外周血单个核细胞中Foxp3转录因子的相对表达量,应用酶联免疫吸附试验检测外周血血浆中Treg相关细胞因子IL-35的表达水平。结果 aGVHD阳性组外周血CD4+CD25+Foxp3+Treg细胞百分比、单个核细胞中Foxp3转录因子的相对表达量、血浆中IL-35的表达水平较aGVHD阴性组和正常对照组明显降低(P<0.05),且经有效治疗后各指标显著升高。在aGVHD阳性组中CD4+CD25+Foxp3+Treg细胞百分比、Foxp3的相对表达量与aGVHD的危险度分级呈负相关(P<0.05)。配对样本资料分析显示,aGVHD发生时外周血CD4+CD25+Foxp3+Treg细胞百分比、Foxp3 mRNA的相对表达量与aGVHD发生前1～2周相比明显下降(P<0.05)。结论异基因造血干细胞移植后患者发生aGVHD时CD4+CD25+Foxp3+Treg细胞的百分比、其转录因子的表达量及IL-35水平均发生明显降低,且在治疗后明显升高,可以为临床预测aGVHD的发生发展及预后提供理论依据和实验基础。Objective To research the relationship between regulatory T cells （Tregs）, IL-35 and acute graft-versus-host disease （aGVHD） in clinical allogeneic bone marrow transplantation （allo-BMT） recipients, and conduct a prospective analysis of peripheral blood Tregs and IL-35 in 23 allo-BMT patients. Methods The percentage of CD4＋CD25＋Foxp3＋ Tregs in peripheral CD4＋T cells was detected by flow cytometry. The relative expression of Foxp3 mRNA transcripts in the mononuclear cells was detected by real time polymerase chain reaction （RT-PCR）. And the level of IL-35 in the plasma was got by Enzyme Linked Immunosorbent Assay （ELISA）. Results The percentage of CD4＋CD25＋Foxp3＋ Tregs, the level of Foxp3 mRNA transcripts and the level of IL-35 in the recipients with aGVHD （n=13） were all significantly lower than those in patients without aGVHD （n=10） and the healthy people （n=21） （P〈0.05）. Tregs decreased linearly with increasing grades of GVHD at onset. Tregs at onset of aGVHD predicted the response to the aGVHD treatment. Paired analysis in aGVHD patients comparing Tregs at aGVHD onset to prior time points （7-14 days） showed significantly decreased in the ratio of CD4＋CD25＋Foxp3＋ regulatory T cells and the level of Foxp3 mRNA（P〈0.05）. Conclusion The ratio of CD4＋CD25＋Foxp3＋ Tregs, Foxp3 mRNA relative transcripts and the level of IL-35 were all decreased in the patients with aGVHD which may have an important prognostic value as a biomarker of acute GVHD.

关 键 词：异基因造血干细胞移植 急性移植物抗宿主病 IL-35 调节性T淋巴细胞 FOXP3 

分 类 号：R551.3[医药卫生—血液循环系统疾病]