CAJ | 학술논문

目的探讨Th17细胞在类风湿关节炎(RA)患者外周血中的水平及意义.方法分离RA患者和健康者外周血单个核细胞(PBMC),免疫磁珠阴选CD4~+T细胞,用或不用非特异性刺激剂(A-CD3、A-CD28),然后加佛波酯/离子霉素(PMA/Ion),经固定,透膜处理进行细胞内染色,流式细胞术(FCM)检测CD4~+T细胞内白细胞介素(IL)-17~+/干扰素(IFN)-γ~+、IL-17~+/IL-6~+水平.分组:①健康对照组;②RA病情稳定组;③RA病情活动组.结果免疫磁珠阴选CD4~+T细胞纯度>90%.RA病情活动组IL-17表达水平(1.54±0.41)显著高于RA病情稳定组(0.70±0.21,P<0.01),二者又显著高于健康对照组(0.42±0.12,P<0.01).用A-CD3、A-CD28、IL-23刺激后,CD4~+T细胞IL-17胞内表达水平较无刺激组显著增加(P<0.05).CD4~+T细胞IFN-γ表达水平呈现与IL-17表达相似的特点,而RA患者与健康对照组IL-6表达水平差异无统计学意义.RA患者IL-17胞内表达水平与IFN-γ、IL-6无显著相关.结论 RA患者外周血CD4~+T细胞中存在异常增高的Th17细胞,且其水平与病情活动相关,有望作为判断RA患者病情活动的指标之一.
목적 탐토Th17세포재류풍습관절염(RA)환자외주혈중적수평급의의.방법 분리RA환자화건강자외주혈단개핵세포(PBMC),면역자주음선CD4~+T세포,용혹불용비특이성자격제(A-CD3、A-CD28),연후가불파지/리자매소(PMA/Ion),경고정,투막처리진행세포내염색,류식세포술(FCM)검측CD4~+T세포내백세포개소(IL)-17~+/간우소(IFN)-γ~+、IL-17~+/IL-6~+수평.분조:①건강대조조;②RA병정은정조;③RA병정활동조.결과 면역자주음선CD4~+T세포순도>90%.RA병정활동조IL-17표체수평(1.54±0.41)현저고우RA병정은정조(0.70±0.21,P<0.01),이자우현저고우건강대조조(0.42±0.12,P<0.01).용A-CD3、A-CD28、IL-23자격후,CD4~+T세포IL-17포내표체수평교무자격조현저증가(P<0.05).CD4~+T세포IFN-γ표체수평정현여IL-17표체상사적특점,이RA환자여건강대조조IL-6표체수평차이무통계학의의.RA환자IL-17포내표체수평여IFN-γ、IL-6무현저상관.결론 RA환자외주혈CD4~+T세포중존재이상증고적Th17세포,차기수평여병정활동상관,유망작위판단RA환자병정활동적지표지일.
Objective To study the expression and significance of Th17 cells from peripheral blood of patients with rheumatoid arthritis (RA). Methods Intracelluar flow cytomete detection of IL-17/IFN-γ and IL-17/IL-6 was established using anti-CD3/Anti-CD28/IL-23 as stimulators after isolation of untouched human CD4~+T cells from PBMC. There were three groups in the present study: ①healthy controls group; ② RA stable group; ③RA active group. Results The isolation of untouched human CD4~+T cells from PBMC was effective and its purity was over 90%. The percentage of intracelluar IL-17 in CD4~+ T cells from RA patients was increased significantly. Such percentage in active group (1.54±0.41) was higher than that of stable group (0.70±0.21, P<0.01) and both of them were higher than those of healthy controls (0.42±0.12, P<0.01). Under anti-CD3/Anti-CD28/IL-23 stimulation, the percentage of intracelluar IL-17 was also increased significantly(P<0.01). The porcentage of intracellular IFN-γ was similar to that of IL-17, while that of IL-6 was not significantly different. There is an correlation between IL-17 and IFN-γ or IL-6. Conclusion There is an abnormal expression of IL-17 and IFN-γ in human CD4~+T cells in RA patients, which is related to disease activity . Th17 cells may be used as a new marker for the assessment of RA activity.