Department of Immunology and Transfusion Medicine, Oslo University Hospital Rikshospitalet, Oslo, Norway; Department of Neurology, Oslo University Hospital Rikshospitalet, Oslo, Norway; Institute of Clinical Medicine, University of Oslo, Oslo, Norway.
Eur J Immunol. 2014 Nov;44(11):3439-52. doi: 10.1002/eji.201444662. Epub 2014 Sep 16.
Epstein-Barr virus (EBV) has long been suggested as a pathogen in multiple sclerosis (MS). Here, we used high-throughput sequencing to determine the diversity, compartmentalization, persistence, and EBV-reactivity of the T-cell receptor (TCR) repertoires in MS. TCR-β genes were sequenced in paired samples of cerebrospinal fluid (CSF) and blood from patients with MS and controls with other inflammatory neurological diseases. The TCR repertoires were highly diverse in both compartments and patient groups. Expanded T-cell clones, represented by TCR-β sequences >0.1%, were of different identity in CSF and blood of MS patients, and persisted for more than a year. Reference TCR-β libraries generated from peripheral blood T cells reactive against autologous EBV-transformed B cells were highly enriched for public EBV-specific sequences and were used to quantify EBV-reactive TCR-β sequences in CSF. TCR-β sequences of EBV-reactive CD8+ T cells, including several public EBV-specific sequences, were intrathecally enriched in MS patients only, whereas those of EBV-reactive CD4+ T cells were also enriched in CSF of controls. These data provide evidence for a clonally diverse, yet compartmentalized and persistent, intrathecal T-cell response in MS. The presented strategy links TCR sequence to intrathecal T-cell specificity, demonstrating enrichment of EBV-reactive CD8+ T cells in MS.
爱泼斯坦-巴尔病毒（EBV）长期以来一直被认为是多发性硬化症（MS）的病原体。在这里，我们使用高通量测序来确定 MS 患者脑脊液（CSF）和血液配对样本中 T 细胞受体（TCR）库的多样性、区室化、持久性和 EBV 反应性。从 MS 患者和其他炎症性神经疾病对照的 CSF 和血液中对 TCR-β 基因进行测序。两个隔室和患者组的 TCR 库都具有高度多样性。代表 TCR-β 序列>0.1%的扩展 T 细胞克隆，在 MS 患者的 CSF 和血液中具有不同的同一性，并持续一年以上。从针对自身 EBV 转化 B 细胞反应的外周血 T 细胞生成的参考 TCR-β 文库高度富含公共 EBV 特异性序列，并用于定量 CSF 中的 EBV 反应性 TCR-β 序列。仅在 MS 患者中，包括几个公共 EBV 特异性序列在内的 EBV 反应性 CD8+T 细胞的 TCR-β 序列在鞘内富集，而 EBV 反应性 CD4+T 细胞的 TCR-β 序列也在对照者的 CSF 中富集。这些数据为 MS 中存在克隆多样性但区室化和持久的鞘内 T 细胞反应提供了证据。所提出的策略将 TCR 序列与鞘内 T 细胞特异性联系起来，证明了 EBV 反应性 CD8+T 细胞在 MS 中的富集。
