目的 探讨类风湿关节炎(rheumatoid arthritis,RA)患者髓系来源的抑制细胞(myeloid-derived suppressor cells,MDSC)对自身B细胞增殖的影响,揭示MDSC在RA发病中的作用。方法 健康对照15例,RA患者38例,按28-关节疾病活动度分值(28-joint disease activity score,DAS28)分为高疾病活动度组(DAS28>5.1)、中疾病活动度组(3.2<DAS28≤5.1)和低疾病活动度组(2.6<DAS28≤3.2), 流式细胞术检测其外周血MDSC细胞的比例;同时分选10例高疾病活动度RA患者外周血B细胞和MDSC细胞体外共培养,羟基荧光素二醋酸盐琥珀酰亚胺酯(carboxyfluorescein diacetate succinimidyl ester,CFSE)法检测B细胞的增殖。结果 高疾病活动度RA外周血MDSC比例(7.13%±2.17%)相对于健康成年人(4.79%±1.02%)、中活动度患者(5.35%±1.36%)和低活动度患者(4.72%±1.08%)明显增加(P<0.05),而中、低疾病活动度RA之间及其二者与健康成年人之间差异皆无统计学意义(P>0.05);MDSC比例与患者DAS28分值成正相关(P<0.05),与年龄、病程、类风湿因子(rheumatoid factor,RF)、抗环瓜氨酸多肽(cyclic citrullinated peptide,CCP)抗体等无相关性;MDSC共培养不抑制反而促进B细胞的增殖(P<0.01)。结论 MDSC在高疾病活动度RA患者外周血中明显增加,与疾病活动度呈正相关,并可在体外促进自身B细胞的增殖,提示MDSC可能通过调节B细胞的功能参与RA的疾病进程。
Objective: To investigate the effect of myeloid-derived suppressor cells (MDSC) on pro-liferation of B lymphocytes in rheumatoid arthritis (RA) patients. Methods: The peripheral blood specimens were collected from 15 healthy adults and 38 RA patients who were divided into high disease activity group, medium activity group and low activity group according to their 28-joint disease activity score (DAS28). And the frequencies of MDSC were determined by flow cytometry. Then, B cells and MDSC were isolated by flow cytometry, respectively. B cells were labeled with carboxyfluorescein diacetate succinimidyl ester (CFSE) and then were co-cultured with MDSC in the presence of 3 mg/L anti-CD40 antibody and 10 mg/L CpG, for 3 days. Flow cytometry was performed to investigate the proliferation of B cells. Results: MDSC expanded markedly in high disease activity patients (7.13%±2.17%) compared with medium (5.35%±1.36%) and low disease activity patients (4.72%±1.08%) or healthy controls (4.79%±1.02%) (P<0.05), and there were no statistical differences between healthy controls, medium and low disease activity RA (P>0.05). Moreover, the frequencies of MDSC were positively correlated with the DAS28 (P<0.05). After co-culture, MDSC significantly promoted B cell proliferation (P<0.01). Conclusion: Our studies showed that MDSC expanded obviously in high disease activity RA patients, and their frequencies were positively correlated with the disease activities. Furthermore, MDSC could promote autologous B cell proliferation remarkably in vitro. These findings suggest that MDSC might be involved in RA pathogenesis through regulating B cell functions.
[1] Feldmann M, Brennan FM, Maini RN. Rheumatoid arthritis [J]. Cell, 1996, 85(3): 307-310.
[2] Bax M, Huizinga TW, Toes RE. The pathogenic potential of autoreactive antibodies in rheumatoid arthritis [J]. Semin Immunopathol, 2014, 36(3): 313-325.
[3] Gabrilovich DI, Nagaraj S. Myeloid-derived suppressor cells as regulators of the immune system [J]. Nat Rev Immunol, 2009, 9(3): 162-174.
[4] Marigo I, Dolcetti L, Serafini P, et al. Tumor-induced tolerance and immune suppression by myeloid derived suppressor cells [J]. Immunol Rev, 2008(222): 162-179.
[5] Crook KR, Liu P. Role of myeloid-derived suppressor cells in autoimmune disease [J]. World J Immunol, 2014, 4(1): 26-33.
[6] Guo C, Hu F, Yi H, et al. Myeloid-derived suppressor cells have a proinflammatory role in the pathogenesis of autoimmune arthritis [J]. Ann Rheum Dis, 2016, 75(1): 278-285.
[7] Edwards JC, Szczepanski L, Szechinski J, et al. Efficacy of B-cell-targeted therapy with rituximab in patients with rheumatoid arthritis [J]. N Engl J Med, 2004, 350(25): 2572-2581.
[8] Young MR, Newby M, Wepsic TH. Hematopoiesis and suppressor bone marrow cells in mice bearing large metastatic Lewis lung carcinoma tumors [J]. Cancer Res, 1987, 47(1): 100-106.
[9] 许为民, 龚爱华, 王慧, 等. 类风湿关节炎患者外周血髓系来源的抑制细胞的检测及与Th1/Th2细胞的相关性分析[J]. 江苏大学学报(医学版), 2013, 23(2): 140-143.
[10] Li Y, Tu Z, Qian S, et al. Myeloid-derived suppressor cells as a potential therapy for experimental autoimmune myasthenia gravis [J]. J Immunol, 2014, 193(5): 2127-2134.
[11] Crook KR, Jin M,Weeks MF, et al. Myeloid-derived suppressor cells regulate T cell and B cell responses during autoimmune disease [J]. J Leukoc Biol, 2015, 97(3): 573-582.
