CAJ | 학술논문

目的:研究系统性红斑狼疮(SLE)患者外周血CD3+、CD4~+、CD8~+T细胞亚群的凋亡特点及其相关的凋亡机制.方法:采用三色荧光流式细胞术检测高活动性SLE患者、低/非活动性SLE患者以及正常对照者外周血T细胞亚群的百分比例、T细胞亚群膜表面Fas/FasL的表达率、T细胞亚群早期凋亡(AV+PI-)的情况;采用ABC-ELISA法测定各组SLE患者和正常对照者血清IL-10水平;对10例血清IL-10水平异常升高的高活动性SLE患者进行体外PBMCs培养实验,其中分别加入抗FasL抗体和抗IL-10抗体,48小时常规培养后分别检测PBMCs中T细胞亚群百分比例、T细胞亚群膜表面Fas/FasL的表达率、T细胞亚群早期凋亡的变化.结果:SLE患者外周血T细胞凋亡异常增多,其中以高活动性SLE患者CD4~+T细胞亚群的凋亡尤为显著(P＜0.05),CD4~+T细胞的异常凋亡与CD4~+和CD8~+T细胞膜表面表达增高的Fas/FasL密切相关(P＜0.05);各组SLE患者外周血血清IL-10水平均明显升高(P＜0.01),高活动性SLE患者更为明显,血清IL-10水平升高不仅与SLE主要临床检验指标相关,而且与CD4~+/CD8~+比值减少、CD4~+T细胞高表达FasL相关(P＜0.05);随访研究显示,随着SLE病情好转、稳定,血清IL-10水平下降明显、T细胞亚群Fas/FasL的表达率明显减少、T细胞亚群凋亡逐渐减少,以上变化均在CD4~+T细胞亚群中体现得最为明显(P＜0.05).结论:SLE患者T细胞活动性异常升高,尤其是CD4~+T细胞亚群凋亡增加,是SLE疾病发展的重要病理机制,IL-10作为能诱导T细胞高表达Fas/FasL的重要调节因子,参与了SLE患者CD4~+T细胞凋亡的免疫调节,SLE患者异常增高的IL-10很町能通过Fas/FasL途径促进了T细胞尤其是CD4~+T细胞亚群的凋亡.
목적:연구계통성홍반랑창(SLE)환자외주혈CD3+、CD4~+、CD8~+T세포아군적조망특점급기상관적조망궤제.방법:채용삼색형광류식세포술검측고활동성SLE환자、저/비활동성SLE환자이급정상대조자외주혈T세포아군적백분비례、T세포아군막표면Fas/FasL적표체솔、T세포아군조기조망(AV+PI-)적정황;채용ABC-ELISA법측정각조SLE환자화정상대조자혈청IL-10수평;대10례혈청IL-10수평이상승고적고활동성SLE환자진행체외PBMCs배양실험,기중분별가입항FasL항체화항IL-10항체,48소시상규배양후분별검측PBMCs중T세포아군백분비례、T세포아군막표면Fas/FasL적표체솔、T세포아군조기조망적변화.결과:SLE환자외주혈T세포조망이상증다,기중이고활동성SLE환자CD4~+T세포아군적조망우위현저(P＜0.05),CD4~+T세포적이상조망여CD4~+화CD8~+T세포막표면표체증고적Fas/FasL밀절상관(P＜0.05);각조SLE환자외주혈혈청IL-10수평균명현승고(P＜0.01),고활동성SLE환자경위명현,혈청IL-10수평승고불부여SLE주요림상검험지표상관,이차여CD4~+/CD8~+비치감소、CD4~+T세포고표체FasL상관(P＜0.05);수방연구현시,수착SLE병정호전、은정,혈청IL-10수평하강명현、T세포아군Fas/FasL적표체솔명현감소、T세포아군조망축점감소,이상변화균재CD4~+T세포아군중체현득최위명현(P＜0.05).결론:SLE환자T세포활동성이상승고,우기시CD4~+T세포아군조망증가,시SLE질병발전적중요병리궤제,IL-10작위능유도T세포고표체Fas/FasL적중요조절인자,삼여료SLE환자CD4~+T세포조망적면역조절,SLE환자이상증고적IL-10흔정능통과Fas/FasL도경촉진료T세포우기시CD4~+T세포아군적조망.
Objective: To study the apoptosis of T lymphocyte subsets of peripheral blood in systemic lupus erythematosus(SLE)and the correlated pathogenesis.Methods: The percentages of T lymphocyte subsets,the positive expression rates of Fas/FasL on T lymphocyte subsets,and the apoptosis of T lymphocyte subsets were determined by three-colorflow cytometry.The level of serum IL-10 was analyzed by ABCELISA.IL-10 antibody and FasL antibody were added respectively into the 48 hours' culture of 10 SLE patients' PBMCs in vitro,whose serum IL-10 significantly increased, and then, the changes of T lymphocyte subsets of SLE PBMCs were determined by three-colodlow cytometry.Results: 1)In SLE, mainly in hyperactive SLE, the apoptesis of CD4~+ T lymphocytes increased significantly(P＜0.05),and the ratio of CD4~+/CD8~+ decreased compared with normal controls,and the abnormal apoptesis of CD4~+ T lymphocytes had a positive correlation with the increased expression rates of Fas/FasL on CD4~+ and CD8~+ T lymphocytes(P＜0.05).2)The serum IL-10 level increased significantly in SLE (P＜0.01), expecially in hyperactive SLE, and serum IL-10 had a positive correlation with the level of serum anti-dsDNA antibodies, CD4~+/CD8 + ratio, and the positive expression rate of FasL on CD4 * T lymphocytes(P＜0.05).3)According to a tracking experiment on two different groups of SLE patients,we found that:with the stabilization of the illness,the level of serum IL-10 decreased significantly, the positive expression rate of Fas/FasL on T lymphocytes,particularly on CD4~+ T lymphocytes, reduced evidently,and the apoptosis of CD4~+ T lymphocytes decreased(P＜0.05).Conclusion: The abnormal activities of T lymphocytes, particularly of CD4~+ T lymphocytes from patients with SLE,which lead to AICD,cause the percentage of CD4~+ T lymphocytes and the ratio of CD4~+/CD8~+ to decrease,disorder immune response, and accelerate the process of SLE.As a key factor in modulating the positive expression rate of Fas/FasL on CD4~+ T lymphocytes,IL-10 plays an important role in causing SLE.