CAJ | 학술논문

目的测定再生障碍性贫血(AA)患者治疗前后外周血CD4+ CD25+ CD127low调节性T细胞(Treg)的数量及叉头翼状螺旋转录因子(FOXP3)mRNA、Notch1 mRNA的表达水平,探讨Treg在AA发病中的作用及其机制.方法流式细胞术检测29例初发AA患者、14例环孢素(CsA)治疗后恢复期及11例治疗后未恢复期患者外周血中CD4+ CD25+ CD127low T细胞、CD4+ CD25+ T细胞的数量,并与正常对照比较;采用RT-PCR检测FOXP3 mRNA和Notch1 mRNA的表达水平,分析两者相关性.结果AA初发组及治疗后未恢复组患者外周血中活化CD4+ CD25+ T细胞占CD4+ T细胞比例分别为(4.3±0.7)%、(4.2±0.6)%,明显高于正常对照组[(2.4±0.8)%](P＜0.05).CsA治疗后恢复组患者比例下降为(2.6±0.7)%(P＜0.05),与对照组比较差异无统计学意义.AA初发组及未恢复组CD4+ CD25+ CD127low T细胞在CD4+ T细胞中的比例分别为(2.4±1.2)%、(2.5±1.1)%,较正常对照组[(7.1±2.7)%]及恢复组[(5.3±1.0)%]明显降低(P值均＜0.01);但后两组比较差异无统计学意义.AA初发组患者FOXP3 mRNA及Notch1 mRNA分别为(0.260±0.011)和(0.018±0.005),较正常对照[(1.307±0.011)和(0.308±0.028)]表达明显下调(P值均＜0.01),治疗后分别为(1.287±0.012)和(0.281±0.013),表达较初发组显著提高(P值均＜0.01),与对照组比较差异无统计学意义(P值均＞0.05).AA患者CD4+ CD25+ CD127low T细胞、FOXP3均与Notch1表达呈正相关性(P值均＜0.01).结论AA患者外周血CD4+ CD25+ CD127low Treg减少,其抑制作用减弱,导致自身反应性T细胞过度活化,抑制造血.其作用机制之一可能与靶细胞表面Notch1分子表达降低相关.
목적 측정재생장애성빈혈(AA)환자치료전후외주혈CD4+ CD25+ CD127low조절성T세포(Treg)적수량급차두익상라선전록인자(FOXP3)mRNA、Notch1 mRNA적표체수평,탐토Treg재AA발병중적작용급기궤제.방법 류식세포술검측29례초발AA환자、14례배포소(CsA)치료후회복기급11례치료후미회복기환자외주혈중CD4+ CD25+ CD127low T세포、CD4+ CD25+ T세포적수량,병여정상대조비교;채용RT-PCR검측FOXP3 mRNA화Notch1 mRNA적표체수평,분석량자상관성.결과AA초발조급치료후미회복조환자외주혈중활화CD4+ CD25+ T세포점CD4+ T세포비례분별위(4.3±0.7)%、(4.2±0.6)%,명현고우정상대조조[(2.4±0.8)%](P＜0.05).CsA치료후회복조환자비례하강위(2.6±0.7)%(P＜0.05),여대조조비교차이무통계학의의.AA초발조급미회복조CD4+ CD25+ CD127low T세포재CD4+ T세포중적비례분별위(2.4±1.2)%、(2.5±1.1)%,교정상대조조[(7.1±2.7)%]급회복조[(5.3±1.0)%]명현강저(P치균＜0.01);단후량조비교차이무통계학의의.AA초발조환자FOXP3 mRNA급Notch1 mRNA분별위(0.260±0.011)화(0.018±0.005),교정상대조[(1.307±0.011)화(0.308±0.028)]표체명현하조(P치균＜0.01),치료후분별위(1.287±0.012)화(0.281±0.013),표체교초발조현저제고(P치균＜0.01),여대조조비교차이무통계학의의(P치균＞0.05).AA환자CD4+ CD25+ CD127low T세포、FOXP3균여Notch1표체정정상관성(P치균＜0.01).결론AA환자외주혈CD4+ CD25+ CD127low Treg감소,기억제작용감약,도치자신반응성T세포과도활화,억제조혈.기작용궤제지일가능여파세포표면Notch1분자표체강저상관.
Objective To quantify the CD4+ CD25+ CD127low regulatory T cell (Treg), the expression levels of forkhead/winged helix transcription factor FOXP3 and Notch1 mRNA in aplastic anemia(AA)patients before and after treatment, and explore the significance of Treg in pathogenesis of AA. Method CD4+ CD25+ and CD4+ CD25+ CD127low T cells in peripheral blood were examined with FACS in 29 AA patients at active phase, 14 at recovery phase, 11 at unrecovery phase, and 15 normal controls. The levels of FOXP3 mRNA and Notch1 mRNA expression were detected with RT-PCR, and the correlations between Treg,FOXP3 mRNA and Notch1 mRNA were analyzed. Results The percentages of peripheral activated CD4+ CD25+ T cells in AA patients at active phase (4.3±0.7)% and unrecovery phase (4.2±0.6)% were significantly higher than those in normal controls (2.4±0.8)% (P＜0.05). The proportion of these cells in AA patients at recovery phase was reduced to (2.6±0.7 )%( P＜0.05 ), being no difference from that in control group. The number of CD4+ CD25+ CD127low T cells in AA patients at active phase (2.4±1.2)% and unrecovery phase (2.5±1.1 )% was decreased significantly compared with those in normal controls (7.1±2.7)% (P＜0.01) and in AA patients at recovery phase (5.3±1.0)% (P＜0.01), there was no difference between the latter two groups. In active phase AA patients, the levels of FOXP3 mRNA and Notch1 mRNA (0.260±0.011 and 0. 018±0.005, respectively) were lower than that in control group (1. 307±0. 011 and 0.308±0. 028, respectively) ( P＜0. 01 and P＜0.01 ). After treatment, the levels significantly increased to 1. 287±0.012 and 0. 281±0. 013 (P＜0. 01 and P＜0. 01), but there was no difference with that of normal controls(P＞0.05). CD4+ CD25+ CD27low T cells and FOXP3 were positively related with Notch1(P＜0. 01) in AA patients. Conclusion The decreased number and suppressive activity of CD4+ CD25+ CD127low Treg cells in the peripheral blood of AA patients cause over-activation of autoreactive T cells and suppression of haematopoiesis. One of the mechanisms maybe the reduced expression of Notch1 in the target cells.