CD127用于特异性识别调节性T淋巴细胞的方法建立及临床应用评价  

CD127, a new biomarker of regulatory T cells：the method and the clinical application

作　　者：袁向亮[1] 沈定丰[2] 卢剑[1] 董平[2] 王剑[1] 李美星[1] 沈立松[1] 

机构地区：[1]上海交通大学医学院附属新华医院检验科,200092 [2]上海交通大学医学院附属新华医院普外科,200092

出　　处：《中华检验医学杂志》2008年第5期499-503,共5页Chinese Journal of Laboratory Medicine

基　　金：上海市教育委员会自然科学基金资助项目（06BZ050）

摘　　要：目的建立用膜表面标志CD4cD妒cD劣界定调节性T淋巴细胞的方法，并评价其临床应用意义。方法五色流式细胞术以Foxp3-FITC／CD127-PE／CD4-PerCP／CD25-APC／CD3-PC7抗体组合鉴定调节性T淋巴细胞，分选CD4CD25^highCD127^low细胞鉴定Foxp3mRNA和蛋白水平的表达情况，比较CD127^low与Foxp3＋界定调节性T淋巴细胞的相关性。用该膜表面标志分析20名健康对照者外周血及35例胃癌患者不同部位CD4cD去调节性T淋巴细胞（Treg）细胞的分布特点及意义。结果用CD4CD25^highCD127^low界定调节性T淋巴细胞的Foxp3蛋白表达率为87．1％，Foxp3mRNA高表达。用膜表面抗原CD4CD25^highCD127^low界定调节性T淋巴细胞与CD4CD。sighFoxp3’具有相关性（r=0．985，P〈0．01）。胃癌患者外周血CD4CD25^highCD127^low调节性T淋巴细胞占CD4T淋巴细胞的比例为（8．67±3．52）％，高于健康对照组[（5．12±2．46）％，t=2．542，P〈0．05]。胃癌患者腹腔积液、肿瘤浸润性淋巴细胞、癌旁淋巴结中CD4CD25^highCD127^low调节性T淋巴细胞占CD4T淋巴细胞比例均高于外周血（分别为t=2．357，P〈0．05；t=6．174，P〈0．01；t=5．481，P〈0．01）。胃癌Ⅰ期＋Ⅱ期患者外周血CD4CD25^highCD127^low细胞占CD4T淋巴细胞的比例为（6．04±2．31）％，Ⅲ期＋Ⅳ期患者为（10．16±2．29）％，其分期差异具有显著统计学意义（t=2．473，P〈0．05）。结论CD4CD。highCD矗≯抗体组合可用于鉴定调节性T淋巴细胞。胃癌患者CD4CD25^highCD127^low调节性T淋巴细胞的比例升高，且增高程度与肿瘤临床分期相关。Objective The aim of the study was to establish the method using CD127 as the new biomarker to identify regulatory T cells （Treg cells） and apply the CD127 to detect the Treg cells in patients with gastric cancer. Methods The phenotypes of Treg cells were analyzed using five-color flow cytometry method Foxp3-FITC/CD127-PE/CD4-PerCP/CD25-APC/CD3-PC7. The mRNA and protein expression of Foxp3 in isolated CD4CD25^highCD127^low Treg cells were detected. The relationships between Foxp3 and CD127 protein expression in CD4＋ T cells from adult human peripheral blood were investigated. PBMCs, Ascites, tumor-infiltration lymphocyte and tumor-draining lymph nodes in 35 patients with gastric cancer and PBMCs in 20 normal healthy donors were evaluated for the proportion of Treg cells, as well as the percentage of the total CD4＋ cells. Results CD4CD25^highCD127^low cells expressed the high Foxp3 in protein level （87.1%） and mRNA level. Within the CD4＋ CD25＋ population, there was a significant correlation between Foxp3 and the CD127^low phenotype （ r = 0. 985, P 〈 0.01 ）. Compared with healthy volunteers, patients with gastric malignancies had a higher proportion of CD4CD25^highCD127^low cells in peripheral blood （t = 2. 542, P 〈 0. 05 ）. The percentages of Treg cells were more abundant in ascites （ t = 2. 357, P 〈 0. 05 ）, TIL （ t = 6. 174, P 〈 0. 01 ） and tumor-draining lymph nodes （ t = 5. 481, P 〈 0. 01 ） of individuals with gastric cancer than that in their blood. There were significant differences in the prevalence of Treg cells between the early and advanced disease stages in gastric cancer [（6.04 ±2.31）% in stage Ⅰ ＋ Ⅱ vs （10. 16 ±2.29）% Ⅲ ＋ Ⅳ,t = 2. 473, P 〈 0. 05 ]. Conclusions The CDI27 biomarker can be used to selectively enrich human Treg cells for in vitro functional studies. The populations of CD4CD25^highCD127^low Treg cells increased with tumor stage in individuals with gastric cancer.

关 键 词：胃肿瘤 受体 白细胞介素7 T淋巴细胞 DNA结合蛋白质类 肿瘤分期 流式细胞术 

分 类 号：R686[医药卫生—骨科学]