上海交通大学学报(医学版) ›› 2019, Vol. 39 ›› Issue (8): 873-.doi: 10.3969/j.issn.1674-8115.2019.08.011

• 论著·基础研究 • 上一篇    下一篇

类风湿关节炎患者 B细胞亚群分析及表没食子儿茶素没食子 酸酯对 B细胞亚群的影响

陈芳倩 1,阎羽欣 1,毛梦寒 1,彭昊 1,金书欣 1,蔡强 2,杨阳 2,岳涛 2,朱琦 2,席晔斌 1,陈广洁 1   

  1. 1. 上海交通大学基础医学院免疫学与微生物学系,上海 200025;2. 上海中医药大学附属光华医院关节内科,上海 200052
  • 出版日期:2019-08-28 发布日期:2019-09-23
  • 通讯作者: 陈广洁,电子信箱:guangjie_chen@163.com。
  • 作者简介:陈芳倩(1996—),女,本科生;电子信箱: fq_chen@foxmail.com。
  • 基金资助:
    国家自然科学基金(81373208,81771731);上海交通大学医学院 2015级以研究为基础的学习项目;上海市大学生创新训练计划(1117008)

Analysis of B cell subsets in rhumatoid arthritis patients and the effect of epigallocatechingallate on B cell subsets

CHEN Fang-qian1, YAN Yu-xin1, MAO Meng-han1, PENG Hao1, JIN Shu-xin1, CAI Qiang2, YANG Yang2, YUE Tao2,ZHU Qi2,XI Ye-bin1, CHEN Guang-jie1   

  1. 1. Department of Immunology and Microbiology, Shanghai Jiao Tong University College of Basic Medical Sciences, Shanghai 200025, China; 2. Department of Arthrology, Guanghua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 200052, China
  • Online:2019-08-28 Published:2019-09-23
  • Supported by:
    National Natural Science Foundation of China, 81373208, 81771731; Shanghai Jiao Tong University School of Medicine Research-based Learning(RBL) Project for Grade 2015; Shanghai University Student Innovation Training Program, 1117008

摘要: 目的 ·探讨类风湿关节炎( rhumatoid arthritis,RA)患者 B细胞亚群特点及表没食子儿茶素没食子酸酯( epigallocatechingallate, EGCG)对 RA患者 B细胞亚群的调节作用。方法 ·纳入年龄、性别匹配的 RA患者和健康对照各 29例,运用配对 t检验分析 2组外周血中 B细胞亚群的差异;根据 28关节疾病活动评分( disease activity score in 28 joints,DAS28)的值将 RA患者分为活动组(2.6 ≤ DAS28<5.1)和高度活动组( DAS28 ≥ 5.1),运用 t检验分析 2组外周血中 B细胞亚群的差异。在 0、10、100 μmol/L EGCG和 2.5 μg/L葡萄球菌 A蛋白共刺激条件下,体外培养 RA患者外周血单个核细胞, 24 h后应用实时聚合酶链反应检测 B细胞活化因子受体(B-cell-activating factor receptor,BAFF-R)的表达水平,48 h后用流式细胞术检测 B细胞亚群。结果 · RA患者的总 B细胞、未分化 B细胞、记忆 B细胞和浆母细胞占淋巴细胞比例和数量均高于健康对照 (P<0.05),而 CD19+ IL-10+调节性 B细胞( regulatory B cell,Breg)占淋巴细胞比例和数量与健康对照相比差异无统计学意义( P>0.05)。10例活动组与 19例高度活动组 RA患者的总 B细胞及各 B细胞亚群(除 CD19+ IL-10+ Breg外)占淋巴细胞比例和数量的差异无统计学意义( P>0.05),6例活动组与 12例高度活动组 RA患者 CD19+ IL-10+ Breg占淋巴细胞比例和数量的差异也无统计学意义(P>0.05)。RA患者外周血总 B细胞所占比例与 IgG型类风湿因子呈弱正相关(r0.308)。EGCG能显著提高 CD19+ IL-10+ Breg占淋巴细胞比例 (P<0.05),且 100 μmol/L EGCG能显著降低总 B细胞中 BAFF-R mRNA的表达水平 (P0.000);但其对未分化 B细胞、记忆 B细胞和浆母细胞占淋巴细胞比例无显著影响( P>0.05)。结论 · B细胞可能在 RA发病中发挥辅助作用,RA患者 CD19+ IL-10+ Breg数量反馈性增加,EGCG可促进 Breg增殖,降低 BAFF-R mRNA表达水平。

关键词: 类风湿关节炎, 表没食子儿茶素没食子酸酯, B细胞亚群, 调节性 B细胞, B细胞活化因子受体

Abstract: Objective · To explore the characteristics of B cell subsets in rheumatoid arthritis (RA) patients and the regulation of epigallocatechingallate (EGCG) on B cell subsets in RA patients. Methods · Twenty-nine age- and sex-matched RA patients and 29 healthy controls were selected, and the difference of B cell subsets in peripheral blood between the two groups was analyzedpaired t-test. According to the value of disease activity score in 28 joints (DAS28), RA patients were divided into active group (2.6≤DAS28<5.1) and highly active group (DAS28≥5.1). The differences of B cell subsets in peripheral blood between the two groups were analyzedt test. Peripheral blood mononuclear cells were cultured in vitro under co-stimulation of 0, 10, 100 μmol/L EGCG and 2.5 μg/L staphylococcal protein A. The level of B-cell-activating factor receptor (BAFF-R) mRNA was detectedreal-time PCR after 24 h, and B cell subsets were detectedflow cytometry after 48 h. Results · The numbers and the proportions of total B cells, undifferentiated B cells,memory B cells and plasmablasts in lymphocytes of RApatients were significantly higher than those of healthy controls (P<0.05), which of CD19+ IL-10+ regulatory B cells (Breg) of RA patients were not significantly different those of healthy controls (P>0.05). There was no significant difference in the numbers and the proportions of total B cells and B cell subsets (except CD19+ IL-10+ Breg) between 10 RA patients of active group and 19 RA patients of highly active group (P>0.05). There was no significant difference in the number and the proportion of CD19+ IL-10+ Breg in lymphocytes between 6 RA patients of active group and 12 RA patients of highly active group (P>0.05). The proportion of total B cells was weakly positively correlated with IgG type rheumatoid factor (r0.308). EGCG could significantly increase the proportion of CD19+ IL-10+ Breg (P<0.05) and 100 μmol/L EGCG could significantly suppress the of BAFF-R mRNA in B cells (P0.000). However, it had no significant effect on the proportions of undifferentiated B cells, memory B cells and plasmablasts in lymphocytes (P>0.05). Conclusion · B cells may play an auxiliary role in the development of RA. The number of CD19+ IL-10+ Breg in RA patients increases as a feedback. EGCG can promote Breg proliferation and suppress BAFF-R mRNA .

Key words: rheumatoid arthritis (RA), epigallocatechingallate (EGCG), B cell subsets, regulatory B cell (Breg), B-cell-activating factor receptor (BAFF-R)

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