类风湿关节炎先天淋巴样细胞的表型特征

O. Boeva, M. T. Berishvili, A. Sizikov, E. Pashkina
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摘要

目前,类风湿性关节炎(RA)被认为是世界范围内最常见的自身免疫性疾病之一,与进行性残疾、系统性器官和组织病变以及国家的社会和经济损失有关。先天淋巴样细胞(innate lymphoid cells, ILS)的研究对于研究RA自身免疫性炎症的发生,特别是细胞的可塑性问题具有现实和重要意义。ILC代表具有功能多样性的组织常驻淋巴样细胞,如T细胞。此外,ILC还通过细胞因子的产生调节免疫应答的定向。少量的白细胞介素存在于血液中,可能是为了迁移到目标器官和组织。因此,研究ILC在RA中的作用将促进对RA发病机制的认识。未来也有可能根据对免疫平衡的影响开发新的治疗策略,以及减少RA的炎症过程。本研究的目的是确定类风湿关节炎中ILCs的亚群组成和表型特征。我们从7例RA患者和6例健康供者中分离和研究外周血单个核细胞(PBMC)。用荧光染料偶联的单克隆抗体对分离的血液MNCs进行染色:谱系特异性(CD3/14/16/19/20/56)和抗fcer1 α - fitc、抗cd274 - apc /Cy7、抗cd127 - percp /Cy5.5、抗cd336 - pe、抗cd117 - apc。ILCs定义为Lin-CD127+。在一般人群中估计CD294+ILCs (ILC2)的数量,将CD117-CD294-ILCs定义为ILC1,将CD117+CD294-ILCs定义为ILC3。用FACS Canto II流式细胞仪(BD Biosciences, USA)分析细胞表型。我们确定了总血液跨国公司中不同ILC亚群(ILC1, ILC2和ILC3)的相对数量。研究表明,与健康供者相比,RA患者的ILC2细胞数量有统计学意义上的显著减少,而供者和患者之间ILC1和ILC3的百分比无显著差异。我们还评估了c-Kit+ILC2的数量;在供者和患者之间,这些细胞的比例没有显著差异。ILCs代表了一群参与RA发病机制的细胞。ILC2在RA中的作用可能是保护性的。ILC失衡可能导致RA的发生。为了更好地了解RA的发病机制,需要进一步研究ILC在该疾病中的亚群特征、表型和功能特征。
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Phenotypic features of innate lymphoid cells in rheumatoid arthritis
Currently, rheumatoid arthritis (RA) is considered among the most common autoimmune diseases worldwide, being associated with progressing disability, systemic organ and tissue lesions, as well as social and economic losses for the state. Studies of innate lymphoid cells (ILS) seems to be actual and significant when studying development of autoimmune inflammation in RA, in particular, the issues of the cell plasticity. ILC represent tissue resident lymphoid cells that display functional diversity, like as T cells. Moreover, ILC regulate orientation of immune response by means of cytokine production. Small amounts of ILCs are present in the bloodstream, presumably for migration to target organs and tissues. Accordingly, the study of ILC in RA will promote understanding of the RA pathogenesis. It is also possible in the future to develop new therapeutic strategies based on the impact on the immunological balance, as well as reducing the inflammatory process in RA. The aim of this study was to determine the subpopulation composition and phenotypic features of ILCs in RA. We have isolated and studied peripheral blood mononuclear cells (PBMC) from 7 patients with RA and 6 healthy donors. The isolated blood MNCs were stained with monoclonal antibodies conjugated to fluorochromes: lineage-specific (CD3/14/16/19/20/56) and anti-FceR1 alpha-FITC, anti-CD294-APC/Cy7, anti-CD127-PerCP/Cy5.5, anti-CD336-PE, anti-CD117-APC. ILCs were defined as Lin-CD127+. The numbers of CD294+ILCs (ILC2) were estimated in the general population, CD117-CD294-ILCs were defined as ILC1, and CD117+CD294-ILCs, as ILC3. The cell phenotype was analyzed with a FACS Canto II flow cytometer (BD Biosciences, USA). We determined relative numbers of different ILC subpopulations (ILC1, ILC2, and ILC3) among the total blood MNCs. It was shown, that the number of ILC2 cells in RA patients was statistically significantly reduced compared to healthy donors, whereas no significant differences in percentage of ILC1 and ILC3 were revealed between donors and patients. We also evaluated the amount of c-Kit+ILC2; there were no significant differences in the proportion of these cells between donors and patients. ILCs represent a population of cells that contribute to the RA pathogenesis. The role of ILC2 in RA is, presumably, protective. The ILC imbalance may contribute to the development of RA. For a better understanding of the RA pathogenesis, further studies of the subpopulation profile, phenotypic and functional characteristics of ILC are required in this disorder.
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