Hunter Patricia J, Nistala Kiran, Jina Nipurna, Eddaoudi Ayad, Thomson Wendy, Hubank Mike, Wedderburn Lucy R
Rheumatology Unit, University College London Institute of Child Health, 30 Guilford Street, London WC1N 1EH, UK.
Arthritis Rheum. 2010 Mar;62(3):896-907. doi: 10.1002/art.27284.
To identify biomarkers in the first synovial fluid (SF) aspirate obtained from children with oligoarticular juvenile idiopathic arthritis (JIA), which could be used to identify children whose disease is likely to extend to a more severe phenotype.
Patients with recent-onset oligoarticular JIA were identified and grouped according to those whose mild disease persisted (persistent disease) or those whose disease would extend from a mild to more severe phenotype (extended-to-be disease) at 1 year after diagnosis. Flow cytometry was used to delineate differences in the mononuclear cell populations between the first blood sample and first SF aspirate from the same patient and between outcome (persistent versus extended-to-be) groups. Proportions of lymphocytes in the joint were modeled on chemotaxis of lymphocytes to CCL5, using Transwell migration assays. Levels of CCL5 in the SF were quantified by enzyme-linked immunosorbent assay. RNA profiles of SF mononuclear cells were compared between groups using the Affymetrix GeneChip hybridization protocol and hierarchical clustering analyses.
Compared with peripheral blood mononuclear cells, SF mononuclear cells displayed an expansion of CD8+ T cells, reduced proportion of B cells, and expansion of CD16- natural killer cells. The lower CD4:CD8 ratio in the SF was recapitulated in vitro by the observed migration of blood T cells in response to CCL5. Synovial CCL5 levels were higher in children whose disease extended to a more severe phenotype. The CD4:CD8 ratio in the SF was significantly lower in patients with extended-to-be oligoarticular JIA (0.57 compared with 0.90 in the persistent disease group, difference 0.33, 95% confidence interval 0.04-0.62; P = 0.009). Gene expression profiling revealed that 344 genes were >1.5-fold differentially expressed between outcome groups (P < 0.05), and these included genes associated with inflammation and macrophage differentiation, which showed increased levels in patients with extended disease at 1 year, and genes associated with immune regulation, which showed increased levels in patients with persistent disease at 1 year.
Analyses of the proportions of synovial lymphocytes, levels of CCL5, and differential gene expression yielded potential biomarkers with which to predict the likelihood of extension of oligoarticular JIA to a more severe disease phenotype.
在少关节型幼年特发性关节炎（JIA）患儿首次滑膜液（SF）抽吸物中鉴定生物标志物，这些标志物可用于识别疾病可能发展为更严重表型的患儿。
确定近期发病的少关节型JIA患者，并根据诊断后1年时疾病轻度持续（持续性疾病）或疾病从轻度发展为更严重表型（发展型疾病）的患者进行分组。采用流式细胞术描绘同一患者的首次血液样本与首次SF抽吸物之间以及结局（持续性与发展型）组之间单核细胞群体的差异。使用Transwell迁移试验，根据淋巴细胞对CCL5的趋化性模拟关节中淋巴细胞的比例。通过酶联免疫吸附测定法定量SF中CCL5的水平。使用Affymetrix GeneChip杂交方案和层次聚类分析比较组间SF单核细胞的RNA谱。
与外周血单核细胞相比，SF单核细胞显示CD8 + T细胞扩增、B细胞比例降低以及CD16 - 自然杀伤细胞扩增。观察到血液T细胞对CCL5的迁移反应，在体外再现了SF中较低的CD4:CD8比值。疾病发展为更严重表型的儿童滑膜CCL5水平较高。发展型少关节型JIA患者SF中的CD4:CD8比值显著低于持续性疾病组（分别为0.57和0.90，差异为0.33，95%置信区间为0.04 - 0.62；P = 0.009）。基因表达谱分析显示，结局组之间有344个基因差异表达>1.5倍（P < 0.05），其中包括与炎症和巨噬细胞分化相关的基因，这些基因在1年时发展型疾病患者中水平升高，以及与免疫调节相关的基因，这些基因在1年时持续性疾病患者中水平升高。
对滑膜淋巴细胞比例、CCL5水平和差异基因表达的分析产生了潜在的生物标志物，可用于预测少关节型JIA发展为更严重疾病表型的可能性。
