北京大学学报(医学版) ›› 2016, Vol. 48 ›› Issue (6): 951-957. doi: 10.3969/j.issn.1671-167X.2016.06.006

• 论著 • 上一篇    下一篇

类风湿关节炎患者外周血滤泡辅助性T细胞水平检测

安乐美,李娟,季兰岚,李光韬,张卓莉△   

  1. (北京大学第一医院风湿免疫科, 北京100034)
  • 出版日期:2016-12-18 发布日期:2016-12-18
  • 通讯作者: 张卓莉 E-mail:zhuoli.zhang@126.com
  • 基金资助:

    国家重点基础研究发展计划(973计划,2010CB529103)资助

Detection of peripheral follicular helper T cells in rheumatoid arthritis

AN Le-mei, LI Juan, JI Lan-lan, LI Guang-tao, ZHANG Zhuo-li△   

  1. (Department of Rheumatology and Clinical Immunology, Peking University First Hospital, Beijing 100034, China)
  • Online:2016-12-18 Published:2016-12-18
  • Contact: ZHANG Zhuo-li E-mail:zhuoli.zhang@126.com
  • Supported by:

    Supported by the Major State Basic Research Development Program of People’s Republic of China (973 Program, 2010CB529103)

摘要:

目的:检测外周血滤泡辅助性T细胞(follicular T helper cells , Tfh cells)的比例及其表面标志,分析与类风湿关节炎(rheumatoid arthritis , RA)患者疾病活动度的关系。方法: 收集40例类风湿关节炎患者及20例正常对照组的外周血,分离外周血单个核细胞(peripheral blood mononuclear cells,  PBMC)及血清,流式细胞术检测PBMC中CD4+CXCR5+Tfh细胞(CXCR5, 趋化因子受体, C-X-C chemokine receptor type 5)的比例及其表面程序性细胞死亡1(programmed death-1, PD-1)、可诱导的协同刺激性因子(inducible co-stimulator, ICOS)、CD40配体(CD40 ligand, CD40L)及白介素21受体(interleukin-21  receptor, IL-21R)的表达情况,实时荧光定量PCR方法检测B 细胞淋巴瘤-6(B-cell lymphoma 6, Bcl-6)、白介素-21(interleukin-21, IL-21)及IL-21R的表达,酶联免疫吸附法(enzyme-linked immunosorbent assay , ELISA)检测血清中IL-21和B 细胞趋化因子CXC配体13 (B cell-attracting chemokine CXC ligand 13, CXCL13)水平。结果: (1)流式细胞术结果提示,与正常对照组比较,RA患者组外周血CXCR5+ CD4+Tfh细胞表达与正常对照组相比明显升高(16.75±3.92  vs. 7.49±1.84,P<0.001);低疾病活动度或缓解组RA外周血CXCR5+ CD4+Tfh细胞表达与正常对照组相比明显升高(16.62±3.43 vs. 7.49±1.84,P<0.001);中疾病活动度组RA外周血CXCR5+ CD4+Tfh细胞表达与正常对照组相比明显升高(16.82±3.07 vs. 7.49±1.84, P<0.001);高疾病活动度组RA外周血CXCR5+ CD4+Tfh细胞表达与正常对照组相比明显升高(16.87±5.50 vs. 7.49±1.84, P<0.001);RA患者外周血ICOS、PD-1、IL-21R在CD4+CXCR5+细胞的表达与正常对照组相比显著升高 (ICOS+ CXCR5+CD4+细胞, 8.37±4.28 vs. 3.72±1.81, P<0.001; PD-1+ CXCR5+CD4+细胞, 1.57±1.10 vs. 0.24±0.30, P=0.035; IL-21R+ CXCR5+CD4+细胞, 4.60±4.05 vs. 0.20±0.19, P=0.006),而CD40L在CD4+CXCR5+细胞的表达在两组间差异无统计学意义(3.38±3.71 vs. 0.54±0.34, P=0.135)。(2)RA患者组较正常对照组外周血中IL-21R mRNA升高(5.00±4.94 vs. 0.74±0.55, P<0.001),而转录因子Bcl-6 mRNA [4.54(3.33, 7.23) vs. 5.31(2.81, 7.44), P=0.329]以及IL-21 mRNA [0.72(0.26, 3.45) vs. 0.56(0.27, 3.71) P=0.195]表达差异无统计学意义(P>0.05);Bcl-6、IL-21和IL-21R mRNA的表达在不同组RA患者之间差异也无统计学意义(P>0.05)。(3)RA患者血清中IL-21、CXCL13水平明显高于正常对照组[IL-21, (200.49±154.56) ng/L vs. (8.21±5.95) ng/L, P<0.001; CXCL13, (43.09±1.28)ng/L vs. (93.72±49.72) ng/L,P<0.001];相关分析显示IL-21、CXCL13浓度与RA疾病活动度正相关(r>0.4),而血清类风湿因子(rheumatoid factor, RF)和抗环瓜氨酸多肽抗体(anti-citrullinated protein antibody, anti-CCP)与RA疾病活动度无相关性(r<0.2)。结论: 外周血Tfh细胞及其相关细胞因子在RA患者中显著升高,说明Tfh细胞可能参与RA的发病机制,阻断Tfh细胞的产生,有望成为治疗RA患者的新的作用靶点。

关键词: 关节炎, 类风湿, T淋巴细胞, 辅助诱导, 细胞因子类, 受体, CXCR5, 白细胞介素21

Abstract:

Objective: To detect cell frequency and surface markers of peripheral CD4+CXCR5+ follicular helper T (Tfh) cells and analyze the correlation between CD4+CXCR5+Tfh cells and rheumatoid arthritis (RA) disease activity. Methods: Forty RA patients meeting the American College of Rheumatology classification criteria for RA and twenty healthy controls (HC) were included. The peripheral blood mononuclear cells and sera were collected. The expressions of CD4+CXCR5+Tfh cells (CXCR5, C-X-C chemokine receptor type 5) and inducible T cell costimulator (ICOS), programmed death 1 positive (PD-1), interleukin-21 receptor (IL-21R) and CD40 ligand (CD40L) positive on CD4+CXCR5+Tfh cells were analyzed by flow cytometry. The transcript levels of B-cell lymphoma 6 (Bcl-6), as well as IL-21 and IL-21R, were measured by real-time polymerase chain reaction. Besides, serum IL-21 and CXCL13 concentrations were determined by enzyme-linked immunosorbent assay. The potential association between Tfh cells and RA disease activity was detected. Results: The cell surface marker of CXCR5+ on CD4+ cells was significantly increasingly higher across the following groups versus HC: total RA patients (16.75±3.92 vs.7.49±1.84, P<0.001); RA patients with low disease activity or remission (16.62±3.43 vs. 7.49±1.84, P<0.001); RA patients with moderate disease activity (16.82±3.07 vs. 7.49±1.84, P<0.001) and RA patients with high disease activity (16.87±5.50 vs. 7.49±1.84, P<0.001). Besides, the percentages of ICOS+, PD-1+, IL-21R+ on CD4+CXCR5+Tfh cells in the RA patients were significantly higher than that of HC (ICOS+CD4+CXCR5+cells, 8.37±4.28 vs. 3.72±1.81, P<0.001; PD-1+CD4+CXCR5+cells, 1.57±1.10 vs. 0.24±0.30, P=0.035; IL-21R+CD4+CXCR5+cells, 4.60±4.05 vs. 0.20±0.19, P=0.006). But the percentage of CD40L+ on CXCR5+CD4+Tfh cells in the RA patients was not significantly higher than that of HC (3.38±3.71 vs. 0.54±0.34, P=0.135). The IL-21R mRNA expression was elevated significantly (5.00±4.94 vs. 0.74±0.55, P<0.001) in the RA patients but not in Bcl-6 mRNA[4.54(3.33, 7.23) vs. 5.31(2.81, 7.44), P=0.329]or IL-21 mRAN[0.72(0.26, 3.45) vs. 0.56(0.27, 3.71), P=0.195]. Additionally, the serum interleukin-21 (IL-21)and CXCL13 levels in the RA patients were higher than in the healthy controls [IL-21, (200.49±154.56) ng/L vs. (8.21±5.95) ng/L, P<0.001; CXCL13, (93.72±49.72) ng/L vs. (43.09±1.28) ng/L, P<0.001] and were both positively correlated with RA disease activity indexes, including erythrocyte sedimentation rate, the disease activity score in 28 joints (ESR-based or CRP-based), clinical disease activity index, and simplified disease activity index. However, none of the Tfh cells, anti-citrullinated protein antibody or rheumatoid factor had any relationship with RA disease activity. Conclusion: Peripheral Tfh cells and their relevant cytokines are higher in RA patients than healthy controls, indicating Tfh cells may participate in the pathogenesis of RA, therefore, blocking the pathway of Tfh cells may be reasonable cellular targets for therapeutic intervention.

Key words: Arthritis, rheumatoid, T-lymphocytes, helper-inducer, Cytokines, Receptors, CXCR5,  , Interleukin-21

中图分类号: 

  • R593.22
[1] 田佳宜,张霞,程功,刘庆红,王世阳,何菁. 系统性红斑狼疮患者血清白细胞介素-2受体α水平及其临床意义[J]. 北京大学学报(医学版), 2021, 53(6): 1083-1087.
[2] 宋志博,耿研,邓雪蓉,张晓慧,张卓莉. 肌肉骨骼超声在指导银屑病关节炎临床分型中的价值[J]. 北京大学学报(医学版), 2021, 53(6): 1061-1066.
[3] 张璐,胡小红,陈澄,蔡月明,王庆文,赵金霞. 类风湿关节炎初治患者颈椎失稳情况及临床特征[J]. 北京大学学报(医学版), 2021, 53(6): 1049-1054.
[4] 邓雪蓉,孙晓莹,张卓莉. 类风湿关节炎患者足踝部体征和超声下病变的一致性[J]. 北京大学学报(医学版), 2021, 53(6): 1037-1042.
[5] 罗靓,霍文岗,张钦,李春. 类风湿关节炎合并角膜溃疡的临床特点和相关因素分析[J]. 北京大学学报(医学版), 2021, 53(6): 1032-1036.
[6] 钟华,徐丽玲,白明欣,苏茵. 类风湿关节炎患者趋化因子CXCL9和CXCL10在骨侵蚀中的作用[J]. 北京大学学报(医学版), 2021, 53(6): 1026-1031.
[7] 娄雪,廖莉,李兴珺,王楠,刘爽,崔若玫,徐健. 类风湿关节炎患者外周血TWEAK基因启动子区甲基化状态及其表达[J]. 北京大学学报(医学版), 2021, 53(6): 1020-1025.
[8] 李正芳,吴雪,武丽君,罗采南,石亚妹,钟岩,陈晓梅,孟新艳. Rhupus综合征的临床特点[J]. 北京大学学报(医学版), 2021, 53(5): 933-937.
[9] 王贵红,左婷,李然,左正才. 瑞巴派特在大鼠痛风性关节炎急性发作中的作用[J]. 北京大学学报(医学版), 2021, 53(4): 716-720.
[10] 马向波,张学武,贾汝琳,高颖,刘洪江,刘玉芳,李英妮. 外周血淋巴细胞亚群检测在系统性硬化症治疗中的应用[J]. 北京大学学报(医学版), 2021, 53(4): 721-727.
[11] 吴俊慧,陈泓伯,武轶群,吴瑶,王紫荆,吴涛,王梦莹,王斯悦,王小文,王伽婷,于欢,胡永华. 2015—2017年北京市2型糖尿病患者骨关节炎患病的相关因素[J]. 北京大学学报(医学版), 2021, 53(3): 518-522.
[12] 赵凯,常志芳,王志华,庞春艳,王永福. 基因沉默肽基精氨酸脱亚胺酶4的表达对胶原诱导关节炎小鼠肺间质病变的影响[J]. 北京大学学报(医学版), 2021, 53(2): 235-239.
[13] 曹迪,王燕,王柳青,孙晓麟,黄妃,孟洋,任丽丽,张学武. 血浆Dickkopf-1在类风湿关节炎患者中的表达及其与外周血T细胞亚群的相关性[J]. 北京大学学报(医学版), 2021, 53(2): 255-260.
[14] 廖栩鹤,王荣福,刘萌,陈雪祺,熊焰,农琳,殷雷,张炳晔,杜毓菁. 18F-FDG PET/CT半定量参数、表皮生长因子受体和间变淋巴瘤激酶基因突变对肺腺癌患者预后评估的价值[J]. 北京大学学报(医学版), 2021, 53(2): 246-254.
[15] 柯岩,张蔷,马云青,李儒军,陶可,桂先革,李克鹏,张洪,林剑浩. 全髋关节置换术治疗脊柱骨骺发育不良患者Tönnis 3级髋关节骨关节炎的早期疗效[J]. 北京大学学报(医学版), 2021, 53(1): 175-182.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
[1] 田增民, 陈涛, Nanbert ZHONG, 李志超, 尹丰, 刘爽. 神经干细胞移植治疗遗传性小脑萎缩的临床研究(英文稿)[J]. 北京大学学报(医学版), 2009, 41(4): 456 -458 .
[2] 郭岩, 谢铮. 用一代人时间弥合差距——健康社会决定因素理论及其国际经验[J]. 北京大学学报(医学版), 2009, 41(2): 125 -128 .
[3] 成刚, 钱振华, 胡军. 艾滋病项目自愿咨询检测的技术效率分析[J]. 北京大学学报(医学版), 2009, 41(2): 135 -140 .
[4] 袁惠燕, 张苑, 范田园. 离子交换型栓塞微球及其载平阳霉素的制备与性质研究[J]. 北京大学学报(医学版), 2009, 41(2): 217 -220 .
[5] 徐莉, 孟焕新, 张立, 陈智滨, 冯向辉, 释栋. 侵袭性牙周炎患者血清中抗牙龈卟啉单胞菌的IgG抗体水平的研究[J]. 北京大学学报(医学版), 2009, 41(1): 52 -55 .
[6] 董稳, 刘瑞昌, 刘克英, 关明, 杨旭东. 氯诺昔康和舒芬太尼用于颌面外科术后自控静脉镇痛的比较[J]. 北京大学学报(医学版), 2009, 41(1): 109 -111 .
[7] 祁琨, 邓芙蓉, 郭新彪. 纳米二氧化钛颗粒对人肺成纤维细胞缝隙连接通讯的影响[J]. 北京大学学报(医学版), 2009, 41(3): 297 -301 .
[8] 李宏亮*, 安卫红*, 赵扬玉, 朱曦. 妊娠合并高脂血症性胰腺炎行血液净化治疗1例[J]. 北京大学学报(医学版), 2009, 41(5): 599 -601 .
[9] 李伟军, 邢晓芳, 曲立科, 孟麟, 寿成超. PRL-3基因C104S位点突变体和CAAX缺失体的构建及表达[J]. 北京大学学报(医学版), 2009, 41(5): 516 -520 .
[10] 丰雷, 王玉凤, 曹庆久. 哌甲酯对注意缺陷多动障碍儿童平衡功能影响的开放性研究[J]. 北京大学学报(医学版), 2007, 39(3): 304 -309 .