CAJ | 학술논문

目的建立用膜表面标志CD4+ CD25high CD-/low127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CD4+CD25highCD-/low127细胞鉴定Foxp3 mRNA和蛋白水平的表达情况,比较CD-/low127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CD4+CD25+调节性T淋巴细胞(Treg)细胞的分布特点及意义.结果用CD4+CD25highCD-/low127界定调节性T淋巴细胞的Foxp3蛋白表达率为87.1%,Foxp3 mRNA高表达.用膜表面抗原CD4+CD25highCD-/low127界定调节性T淋巴细胞与CD4+CD25highFoxp3+具有相关性(r=0.985,P＜0.01).胃癌患者外周血CD25highCD-/low127调节性T淋巴细胞占CD4+T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CD4+CD25highCD-/low127调节性T淋巴细胞占CD4+T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CD25high CD-/low127Treg细胞占CD4+T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 CD4+CD25highCD-/low127抗体组合可用于鉴定调节性T淋巴细胞.胃癌患者CD4+CD25highCD-/low127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.
목적 건립용막표면표지CD4+ CD25high CD-/low127계정조절성T림파세포적방법,병평개기림상응용의의.방법 오색류식세포술이Foxp3-FITC/CD127-PE/CD4-PerCP/CD25-APC/CD3-PC7항체조합감정조절성T림파세포,분선CD4+CD25highCD-/low127세포감정Foxp3 mRNA화단백수평적표체정황,비교CD-/low127여Foxp3+계정조절성T림파세포적상관성.용해막표면표지분석20명건강대조자외주혈급35례위암환자불동부위CD4+CD25+조절성T림파세포(Treg)세포적분포특점급의의.결과 용CD4+CD25highCD-/low127계정조절성T림파세포적Foxp3단백표체솔위87.1%,Foxp3 mRNA고표체.용막표면항원CD4+CD25highCD-/low127계정조절성T림파세포여CD4+CD25highFoxp3+구유상관성(r=0.985,P＜0.01).위암환자외주혈CD25highCD-/low127조절성T림파세포점CD4+T림파세포적비례위(8.67 ±3.52)%,고우건강대조조[(5.12±2.46)%,t=2.542,P＜0.05].위암환자복강적액、종류침윤성림파세포、암방림파결중CD4+CD25highCD-/low127조절성T림파세포점CD4+T림파세포비례균고우외주혈(분별위t=2.357,P＜0.05;t=6.174,P＜0.01;t=5.481,P＜0.01).위암Ⅰ기+Ⅱ기환자외주혈CD25high CD-/low127Treg세포점CD4+T림파세포적비례위(6.04±2.31)%,Ⅲ기+Ⅳ기환자위(10.16±2.29)%,기분기차이구유현저통계학의의(t=2.473,P＜0.05).결론 CD4+CD25highCD-/low127항체조합가용우감정조절성T림파세포.위암환자CD4+CD25highCD-/low127조절성T림파세포적비례승고,차증고정도여종류림상분기상관.
Objective The aim of the study WSfl to establish the method using CDl27 as the new biomarker to identify regulatory T cells(Treg cells)and apply the CD 127 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 CD+4 CD25high CD127-/low Treg cells were detected.The relationships between Foxp3 and CD127 protein expression in CD4+ T cells from aduh human peripheral blood were investigated.PBMCs,Ascltes,turnor-infihration 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 ceils,as well as the percentage ot the total CD +4 cells.Results CD4+ CD25 high CD-127 low ceils expressed the high Foxp3 in protein level(87.1%)and mRNA level.Within the CD4+CD+25 population,there was a significant correlation between Foxp3 and the CD127low phenotype(r=0.985,P<0.01).Compared with healthy olunteers,patients with gastric malignancies had a higher proportion of CD4+4 Cdhigh 25 CD-low127 cells in peripheral blood(t=2.542,P<0.05).The Dereentages 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 ceils between the early and advanced disease stages in gastric cancer[(6.04±2.31)%in stageⅠ+Ⅱ VB(10.16±2.29)% Ⅲ+Ⅳ,t=2.473,P<0.05].Conclusions The CD127 biomarker can be used to selectively enrich human Treg cells for in vitro functional studies.The populations of CD4+ CD high25 CD127-/low Treg cells increased ith tumor stage in individuals with gastric cancer.