CAJ | 학술논문

目的研究系统性红斑狼疮(SEE)外周血中调节性T细胞不同标志以及调节性T细胞在SLE发病中的作用;探讨CD127与Foxp3的相关性,明确CD127定义调节性T细胞的特异性;鉴定CD4~+CD25~+CD127~(low/-)T淋巴细胞免疫抑制功能.方法 ①采用四色直接荧光素标记法和多参数流式细胞术检测40例SLE患者(19例初发和21例缓解)及15名健康对照外周血CD4~+CD25~+T淋巴细胞、CD4~+CD25~+CD127~(low-)T淋巴细胞、CD4~+CD25~+Foxp3~+T淋巴细胞、CD4~+CD25~(high)T淋巴细胞、CD4~+CD25~(high)CD127~(low/-)T淋巴细胞、CD4~+CD25~(high)Foxp3~+T淋巴细胞以及CD4~+CD127~(low/-)Foxp3~+T淋巴细胞占CD4~+T淋巴细胞的比率,并且将7种调节性T细胞比率与外周血抗双链DNA(dsDNA)等抗体及SLE疾病活动指数(SLEDA1)评分等进行相关性分析.②以流式细胞分选术结合细胞培养技术,检测和分析3例SLE患者和4名健康人外周血中CD4~+CD25~+CD127~(low/-)调节性T细胞对CD4~+CD25~-效应性T细胞增殖的抑制作用.采用两样本均数的t检验,重复测量的方差分析,Pearson相关与Spearman相关分析进行统计学处理.结果 ①SLE患者组7种调节性T细胞比率分别为(6.1±1.7)%,(3.1±1.3)%,(2.1±1.0)%,(1.6±0.3)%,(0.97±0.28)%,(0.69±0.23)%和(0.71±0.35)%.与健康对照组比较:SLE患者组前6种调节性T细胞比率均低于健康对照组(P<0.05).②SLE患者组:CD4~+CD25~+Foxp3~+、CD4~+CD25~(high)Foxp3~+T淋巴细胞比率与IgA呈正相关;CD4~+CD25~(high)CD127~(low/-)T淋巴细胞比率与抗SSB抗体呈正相关.③SLE患者初发组和缓解组比较:SLE患者初发组7种调节性T细胞中除CD4~+CD127~(low/-)Foxp3~+T淋巴细胞比率外,其余均低于缓解组(P<0.05).④SLE患者初发组治疗前后比较:激素治疗前6种调节性T细胞比率均低于激素治疗后(P<0.05).⑤SLE患者初发组、缓解组和对照组中,CD4~+CD25~+T淋巴细胞及CD4~+CD25~(high)T淋巴细胞中Foxp3的表达与CD127低表达均呈正相关.⑥SLE患者、健康人CD4+CD25-效应性T细胞的体外增殖都可以被自身CD4~+CD25~+CD127~(low/-)调节性T细胞所抑制,但SLE患者的抑制率明显低于健康对照.结论 SLE的免疫异常可能与调节性T细胞的数量和功能缺陷有关;CD127可能代替Foxp3作为调节性T细胞特异性的表面标记物. 目的研究繫統性紅斑狼瘡(SEE)外週血中調節性T細胞不同標誌以及調節性T細胞在SLE髮病中的作用;探討CD127與Foxp3的相關性,明確CD127定義調節性T細胞的特異性;鑒定CD4~+CD25~+CD127~(low/-)T淋巴細胞免疫抑製功能.方法 ①採用四色直接熒光素標記法和多參數流式細胞術檢測40例SLE患者(19例初髮和21例緩解)及15名健康對照外週血CD4~+CD25~+T淋巴細胞、CD4~+CD25~+CD127~(low-)T淋巴細胞、CD4~+CD25~+Foxp3~+T淋巴細胞、CD4~+CD25~(high)T淋巴細胞、CD4~+CD25~(high)CD127~(low/-)T淋巴細胞、CD4~+CD25~(high)Foxp3~+T淋巴細胞以及CD4~+CD127~(low/-)Foxp3~+T淋巴細胞佔CD4~+T淋巴細胞的比率,併且將7種調節性T細胞比率與外週血抗雙鏈DNA(dsDNA)等抗體及SLE疾病活動指數(SLEDA1)評分等進行相關性分析.②以流式細胞分選術結閤細胞培養技術,檢測和分析3例SLE患者和4名健康人外週血中CD4~+CD25~+CD127~(low/-)調節性T細胞對CD4~+CD25~-效應性T細胞增殖的抑製作用.採用兩樣本均數的t檢驗,重複測量的方差分析,Pearson相關與Spearman相關分析進行統計學處理.結果 ①SLE患者組7種調節性T細胞比率分彆為(6.1±1.7)%,(3.1±1.3)%,(2.1±1.0)%,(1.6±0.3)%,(0.97±0.28)%,(0.69±0.23)%和(0.71±0.35)%.與健康對照組比較:SLE患者組前6種調節性T細胞比率均低于健康對照組(P<0.05).②SLE患者組:CD4~+CD25~+Foxp3~+、CD4~+CD25~(high)Foxp3~+T淋巴細胞比率與IgA呈正相關;CD4~+CD25~(high)CD127~(low/-)T淋巴細胞比率與抗SSB抗體呈正相關.③SLE患者初髮組和緩解組比較:SLE患者初髮組7種調節性T細胞中除CD4~+CD127~(low/-)Foxp3~+T淋巴細胞比率外,其餘均低于緩解組(P<0.05).④SLE患者初髮組治療前後比較:激素治療前6種調節性T細胞比率均低于激素治療後(P<0.05).⑤SLE患者初髮組、緩解組和對照組中,CD4~+CD25~+T淋巴細胞及CD4~+CD25~(high)T淋巴細胞中Foxp3的錶達與CD127低錶達均呈正相關.⑥SLE患者、健康人CD4+CD25-效應性T細胞的體外增殖都可以被自身CD4~+CD25~+CD127~(low/-)調節性T細胞所抑製,但SLE患者的抑製率明顯低于健康對照.結論 SLE的免疫異常可能與調節性T細胞的數量和功能缺陷有關;CD127可能代替Foxp3作為調節性T細胞特異性的錶麵標記物.
목적 연구계통성홍반랑창(SEE)외주혈중조절성T세포불동표지이급조절성T세포재SLE발병중적작용;탐토CD127여Foxp3적상관성,명학CD127정의조절성T세포적특이성;감정CD4~+CD25~+CD127~(low/-)T림파세포면역억제공능.방법 ①채용사색직접형광소표기법화다삼수류식세포술검측40례SLE환자(19례초발화21례완해)급15명건강대조외주혈CD4~+CD25~+T림파세포、CD4~+CD25~+CD127~(low-)T림파세포、CD4~+CD25~+Foxp3~+T림파세포、CD4~+CD25~(high)T림파세포、CD4~+CD25~(high)CD127~(low/-)T림파세포、CD4~+CD25~(high)Foxp3~+T림파세포이급CD4~+CD127~(low/-)Foxp3~+T림파세포점CD4~+T림파세포적비솔,병차장7충조절성T세포비솔여외주혈항쌍련DNA(dsDNA)등항체급SLE질병활동지수(SLEDA1)평분등진행상관성분석.②이류식세포분선술결합세포배양기술,검측화분석3례SLE환자화4명건강인외주혈중CD4~+CD25~+CD127~(low/-)조절성T세포대CD4~+CD25~-효응성T세포증식적억제작용.채용량양본균수적t검험,중복측량적방차분석,Pearson상관여Spearman상관분석진행통계학처리.결과 ①SLE환자조7충조절성T세포비솔분별위(6.1±1.7)%,(3.1±1.3)%,(2.1±1.0)%,(1.6±0.3)%,(0.97±0.28)%,(0.69±0.23)%화(0.71±0.35)%.여건강대조조비교:SLE환자조전6충조절성T세포비솔균저우건강대조조(P<0.05).②SLE환자조:CD4~+CD25~+Foxp3~+、CD4~+CD25~(high)Foxp3~+T림파세포비솔여IgA정정상관;CD4~+CD25~(high)CD127~(low/-)T림파세포비솔여항SSB항체정정상관.③SLE환자초발조화완해조비교:SLE환자초발조7충조절성T세포중제CD4~+CD127~(low/-)Foxp3~+T림파세포비솔외,기여균저우완해조(P<0.05).④SLE환자초발조치료전후비교:격소치료전6충조절성T세포비솔균저우격소치료후(P<0.05).⑤SLE환자초발조、완해조화대조조중,CD4~+CD25~+T림파세포급CD4~+CD25~(high)T림파세포중Foxp3적표체여CD127저표체균정정상관.⑥SLE환자、건강인CD4+CD25-효응성T세포적체외증식도가이피자신CD4~+CD25~+CD127~(low/-)조절성T세포소억제,단SLE환자적억제솔명현저우건강대조.결론 SLE적면역이상가능여조절성T세포적수량화공능결함유관;CD127가능대체Foxp3작위조절성T세포특이성적표면표기물.
Objective To study the role of peripheral blood T regulatory cells (Treg) markers, as well as different Treg cells in the pathogenesis of systemic lupus erythematosus (SLE) and explore the correla-tion betweenCD127 and Foxp3. The immunosuppressive effect of CD4~+CD25~+CD127~(low/-) T cell is explored. Methods ①Four-color direct fluorescence-labeled and multi-parameter flow cytometry were used to detect peripheral blood CD4~+CD25~+ T cells and other Treg cells accounted for the proportion of CD4~+ T cells in 40 SLE patients (19 cases with active disease 21 cases in remission) and 15 healthy controls. Meanwhile, its correlations with anti-dsDNA antibodies among 7 groups were analyzed.②Flow cytometry sorting combined with cell culture were applied to detect and analyze the proliferation inhibition effect of CD4~+CD25~+CD127~(low/-)regulatory T cells to CD4~+CD25~-effector T cell.Two independent samples t test,ANOVA for repeated measures,Pearson's correlation and Spearman's correlation were used for statistical analysis.Results ①The cell ratio of the 7 groups of SLE patients was(6.1±1.7)%,(3.1±1.3)%,(2.1±1.0)%,(1.6±0.3)%(0.97±O.28)%,(0.69±0.23)%and(O.71±0.35)%respectively.In the SLE group,the proportion of the first 6 groups was lower than the control group (P<0.05).For CD4~+CD127~(low/-)Foxp3~+,there wag no difference in the two groups(P>O.05).②CD4~+CD25~+Foxp3~+,CD4~+CD25~(high)Foxp3~+ T cells and IgA ratio Was positivelv correlated.While CD4~+CD25~(high)CD127~(low/-) T cells and anti-SSB antibodies was positively correlated in SLE patients.③The seven group of cells,in addition to CD4~+CDl27~(low/-)Foxp3~+ T cell ratio were lower in early-onset SLE patients than those in patients at remission(P<0.05).④The cell propor-tion of the first 6 groups was lower than that of post steroid treatment(P<0.05).⑤Foxp3 expression of CD4~+CD25~+T ceils and CD4~+CD25~(high) T cells was positively correlated with low expression of CDl27 in the early onset group,remission group and the control group.⑥The CD4~+CD25~- effect T cells could be suppressed by their own CD4~+CD25~+CDl27~(low/-) regulatonry T cells in vitro,and the inhibition of SLE patients was significantly lower than controls.Conclusion The immunological abnormality of SLE may be associated with the qnatity and functional defects of immune regulatory T cells.CDl27 may be a possible alternative to Foxp3 regulatory T cell-specific surface markers.