Department of Clinical Immunology and Rheumatology, Academic Medical Center/ University of Amsterdam, Amsterdam, The Netherlands.
Ann Rheum Dis. 2012 Jun;71(6):1088-93. doi: 10.1136/annrheumdis-2011-200612. Epub 2012 Jan 31.
To profile quantitatively the T-cell repertoire in multiple joints and peripheral blood of patients with recent onset (early) or established rheumatoid arthritis (RA) using a novel next-generation sequencing protocol to identify potential autoreactive clones.
Synovium of patients with recent onset (early) RA (<6 months) (n=6) or established RA (>18 months) (n=6) was screened for T-cell clones by sequencing over 10 000 T-cell receptors (TCR) per sample. T cells from paired blood samples were analysed for comparison. From two patients synovial T cells were obtained from multiple inflamed joints. The degree of expansion of each individual clone was based on its unique CDR3 sequence frequency within a sample. Clones with a frequency of over 0.5% were considered to be highly expanded clones (HEC).
In early RA synovium, the T-cell repertoire was dominated by 35 HEC (median, range 2-70) accounting for 56% of the TCR sequenced. The clonal dominance in the synovium was patient specific and significantly greater than in established RA (median of 11 HEC (range 5-24) in established RA synovium accounting for 9.8% of T cells; p<0.01). 34% (range 28-40%) of the most expanded T-cell clones were shared between different joints in the same patients, compared with only 4% (range 0-8%) between synovium and blood (p=0.01).
In RA, a systemic autoimmune disease, the inflamed synovium forms a niche for specific expanded T-cell clones, especially in early disease. This suggests that, at least in RA, autoreactive T cells should be addressed specifically in the inflamed tissue, preferably in the early phase of the disease.
使用新型下一代测序方案定量描绘早期或已确诊类风湿关节炎（RA）患者多个关节和外周血中的 T 细胞受体（TCR）谱，以鉴定潜在的自身反应性克隆。
对近期（早期）RA（<6 个月）（n=6）或已确诊 RA（>18 个月）（n=6）患者的滑膜进行 TCR 测序，每个样本超过 10000 个 TCR，筛选 T 细胞克隆。对配对血样进行分析以进行比较。从两名患者的多个炎症关节中获取滑膜 T 细胞。根据其在样本中的独特 CDR3 序列频率确定每个克隆的扩增程度。频率超过 0.5%的克隆被认为是高度扩增克隆（HEC）。
在早期 RA 滑膜中，T 细胞受体谱主要由 35 个 HEC（中位数，范围 2-70）组成，占测序 TCR 的 56%。滑膜中的克隆优势具有患者特异性，明显高于已确诊 RA（中位数为 11 个 HEC（范围 5-24），占滑膜 T 细胞的 9.8%；p<0.01）。在同一患者的不同关节之间，34%（范围 28-40%）的最扩增 T 细胞克隆是共享的，而在滑膜和血液之间只有 4%（范围 0-8%）是共享的（p=0.01）。
在 RA 等系统性自身免疫性疾病中，炎症滑膜为特定扩增的 T 细胞克隆形成了一个龛位，尤其是在疾病早期。这表明，至少在 RA 中，自身反应性 T 细胞应在炎症组织中特异性处理，最好在疾病早期阶段。
