CAJ | 학술논문

目的研究异基因造血干细胞移植(allo-HSCT)后CD4+T细胞及CD8+T细胞中CD25+Foxp3+(CD4+Treg细胞或CD8+Treg细胞)、IL-17a(Th17细胞或Tc17细胞)及IFN-γ(Th1细胞或Tc1细胞)表达以及CD4+CD25+CD 127+效应性T细胞(Tcon细胞)重建,比较HLA相合与不合allo-HSCT后T细胞亚群重建的异同.方法收集2011年12月至2012年10月在北京大学血液病研究所进行allo-HSCT后未发生急性移植物抗宿主病的成年血液病患者20例,其中HLA相合10例、HLA不合10例,并留取10份健康供者外周血标本作为健康对照.采用8色流式细胞学检测技术检测CD4+T细胞及CD8+T细胞上CD25Toxp3+、CD 127、IL-17a及IFN-γ表达.结果 ①HLA全合组与不合组患者移植后2个月内CD3+T细胞、CD4+T细胞、CD8+T细胞数量重建两组间差异无统计学意义.②HLA相合组CD4+ Treg细胞[+30 d,8.46(0.36～27.41)细胞数/μl对1.10(0.04～8.03)细胞数/μl,P＜0.05；+60 d,8.50(1.16～36.20)细胞数/μl对2.73(0.34～6.84)细胞数/μl,P＜0.05]、CD4+CD25+CD127+效应性T细胞[+30d,72.69(3.85～211.73)细胞数/μl对13.41(0.48～96.17)细胞数/μl,P＜0.05;+60d,100.85(16.28～267.20)细胞数/μl对47.75(6.34～143.04)细胞数/μl,P＜0.05]、Th17细胞[+30 d,2.34(0.02～6.87)细胞数/μl对0.20(0.02～1.34)细胞数/μl,P＜0.05; +60 d,1.90 (0.36～7.82)细胞数/μl对0.46(0.03～1.39)细胞数/μl,P＜0.05]、Tc17细胞数量(+30 d,1.08(0.07～15.03)细胞数/μl对0.25(0.01～0.81)细胞数/μl,P＜0.05；+60 d,1.85(0.63～26.57)细胞数/μl对0.46(0.01～3.66)细胞数/μl,P＜0.05]在+30 d及+60 d均明显高于HLA 不合组,Th1细胞及Tc1细胞数量重建两组间差异无统计学意义.③Th1与Th17细胞比值及Tc1与Tc17的比值,HLA不合组均高于HLA相合组患者.结论 HLA相合与不合allo-HSCT后T细胞亚群重建存在差异,这种差异有可能部分解释HLA相合与不合allo-HSCT后移植物抗宿主病发病率及累及部位的不同.
목적 연구이기인조혈간세포이식(allo-HSCT)후CD4+T세포급CD8+T세포중CD25+Foxp3+(CD4+Treg세포혹CD8+Treg세포)、IL-17a(Th17세포혹Tc17세포)급IFN-γ(Th1세포혹Tc1세포)표체이급CD4+CD25+CD 127+효응성T세포(Tcon세포)중건,비교HLA상합여불합allo-HSCT후T세포아군중건적이동.방법 수집2011년12월지2012년10월재북경대학혈액병연구소진행allo-HSCT후미발생급성이식물항숙주병적성년혈액병환자20례,기중HLA상합10례、HLA불합10례,병류취10빈건강공자외주혈표본작위건강대조.채용8색류식세포학검측기술검측CD4+T세포급CD8+T세포상CD25Toxp3+、CD 127、IL-17a급IFN-γ표체.결과 ①HLA전합조여불합조환자이식후2개월내CD3+T세포、CD4+T세포、CD8+T세포수량중건량조간차이무통계학의의.②HLA상합조CD4+ Treg세포[+30 d,8.46(0.36～27.41)세포수/μl대1.10(0.04～8.03)세포수/μl,P＜0.05；+60 d,8.50(1.16～36.20)세포수/μl대2.73(0.34～6.84)세포수/μl,P＜0.05]、CD4+CD25+CD127+효응성T세포[+30d,72.69(3.85～211.73)세포수/μl대13.41(0.48～96.17)세포수/μl,P＜0.05;+60d,100.85(16.28～267.20)세포수/μl대47.75(6.34～143.04)세포수/μl,P＜0.05]、Th17세포[+30 d,2.34(0.02～6.87)세포수/μl대0.20(0.02～1.34)세포수/μl,P＜0.05; +60 d,1.90 (0.36～7.82)세포수/μl대0.46(0.03～1.39)세포수/μl,P＜0.05]、Tc17세포수량(+30 d,1.08(0.07～15.03)세포수/μl대0.25(0.01～0.81)세포수/μl,P＜0.05；+60 d,1.85(0.63～26.57)세포수/μl대0.46(0.01～3.66)세포수/μl,P＜0.05]재+30 d급+60 d균명현고우HLA 불합조,Th1세포급Tc1세포수량중건량조간차이무통계학의의.③Th1여Th17세포비치급Tc1여Tc17적비치,HLA불합조균고우HLA상합조환자.결론 HLA상합여불합allo-HSCT후T세포아군중건존재차이,저충차이유가능부분해석HLA상합여불합allo-HSCT후이식물항숙주병발병솔급루급부위적불동.
Objective To compare the differences of the T helper cell reconstitution kinetics beween HLA matched or HLA mismatched allo-HSCT through exploring the reconstitution kinetics of CD4+ CD25+Foxp3+ cells (CD4+Treg),CD8+CD25+Foxp3 + cells (CD8+Treg),CD4+CD25 CD 127+ conventional T cells (Tcon) and the secretion of IL-17a and IFN-r in CD4+ T cells (Th 17 and Th1 cells) or CD8+ T cells (Tc17 and Tc17 cells) post allogeneic hematopoietic stem cells transplantation (allo-HSCT).Methods From December 2011 to October 2012,the peripheral blood (PB) of 20 patients undergoing HLA matched (10 patients) or mismatched (10 patients) allo-HSCT without acute graft-versus-host disease (aGVHD)and of 10 related healthy donors were collected to analyze the expression of CD25+Foxp3+,IL-17a,IFN-γ and CD127 expression through 8-colour Flow cytometer.Results ①The reconstitution kinetics of CD3+ T cells,CD4+ T cells,CD8+ T cells absolute numbers were comparable within 2 month post HLA matched and mismatched transplantation.②The absolute numbers of CD4+ Treg cells [+30 d,8.46 (0.36-27.41) cells/μl 1.10 (0.04-8.03) cells/μl,P＜0.05; + 60 d,8.50 (1.16-36.20) cells/μl vs 2.73 (0.34-6.84) cells/μl,P＜0.05],Tcon cells[+30 d,72.69(3.85-211.73)cells/μl vs 13.41 (0.48-96.17)cells/μl,P＜0.05; +60 d,100.85 (16.28-267.20)cells/μl vs 47.75 (6.34-143.04)cells/μl,P＜0.05],as well as Th17 cells [+30 d,2.34(0.02-6.87) cells/μl vs 0.20 (0.02-1.34)cells/μl,P＜0.05; +60 d,1.90 (0.36-7.82) cells/μl vs 0.46 (0.03-1.39)cells/μl,P＜0.05] and Tcl7 cells [+30 d,1.08 (0.07-15.03) cells/μl vs 0.25 (0.01-0.81) cells/μl,P＜0.05;+60 d,1.85 (0.63-26.57)cells/μl vs 0.46(0.01-3.66)cells/μl,P＜0.05] within 2 month post HLA matched HSCT were significantly higher than those post HLA-mismatched HSCT.However,the absolute numbers of Th1 cells or Tc1 cells within 2 month post HLA-matched or HLA-mismatched HSCT were comparable.③The ratio of Th1 and Th17 cells,or the ratio of Tc1 and Tc17 cells were significantly higher within 2 month post HLA-mismatched allo-HSCT compared to those post HLA-matched HSCT.Conclusion The reconstitution kinetics of T helper cells subset were different at early stage post HLA-matched or HLA-mismatched allo-HSCT,which might be help to explain the different rate or the different involved organ of the acute graft-versus-host diseases (aGVHD) post HLA-matched or-mismatched allo-HSCT.