类风湿关节炎患者外周血Th17细胞的检测及意义  

Expression and significance of Th17 cells from peripheral blood of patients with rheumatoid arthritis

作　　者：芮金兵[1] 李晶[1] 尤海燕[2] 吴玲[1] 裘影影[1] 罗燕萍[1] 宋晓丽[1] 焦志军[2] 

机构地区：[1]江苏大学附属医院风湿科,镇江212001 [2]江苏大学附属医院检验科,镇江212001

出　　处：《中华风湿病学杂志》2009年第11期732-735,共4页Chinese Journal of Rheumatology

基　　金：国家自然科学基金（30872335）;江苏省镇江市重点医学人才资助项目（2005009）

摘　　要：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），然后加佛波酯腐子霉素（PMMIon），经固定／透膜处理进行细胞内染色，流式细胞术（FCM）检测CD4＋T细胞内自细胞介素（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。Objective To study the expression and significance of Thl7 cells from peripheral blood of patients with rheumatoid arthritis （RA）. Methods Intracelluar flow cytometc 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： （1） healthy controls group; （2） RA stable group; （3） RA active group. Results The isolation of untouched human CD4^＋T ceils 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 percentage of intraeellular IFN-~ 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.

关 键 词：关节炎 类风湿 TH17细胞 细胞内染色 

分 类 号：R686[医药卫生—骨科学]