Neuroimmunology Branch, Viral Immunology Section, National Institute of Neurological Disorder and Stroke, NIH, Bethesda, Maryland, United States.
Bioinformatics Section, National Institute of Neurological Disorder and Stroke, NIH, Bethesda, Maryland, United States.
Sci Rep. 2019 Jan 23;9(1):344. doi: 10.1038/s41598-018-36274-7.
In this study we characterized the TCR repertoire profiles in patients with chronic progressive inflammatory neurological disorders including HAM/TSP, associated with human T-cell lymphotropic virus type I (HTLV-I) infection, and multiple sclerosis (MS), an inflammatory, demyelinating disease of the CNS of unknown etiology. We hypothesized that a T-cell receptor (TCR) clonal repertoire 'signature' could distinguish HAM/TSP patients from healthy controls, as well as from patients with a more heterogeneous CNS-reactive inflammatory disease such as MS. In this study, we applied an unbiased molecular technique - unique molecular identifier (UMI) library-based strategy to investigate with high accuracy the TCR clonal repertoire by high throughput sequencing (HTS) technology. cDNA-TCR β-chain libraries were sequenced from 2 million peripheral mononuclear cells (PBMCs) in 14 HAM/TSP patients, 34 MS patients and 20 healthy controls (HC). While HAM/TSP patients showed a higher clonal T-cell expansion compared to MS and HC, increase of the TCR clonal expansion was inversely correlated with the diversity of TCR repertoire in all subjects. In addition, longitudinal analysis of TCR repertoires from HAM/TSP patients demonstrated a correlation of the TCR clonal expansion with HTLV-I proviral load. Surprisingly, MS patients showed a higher diversity of TCR repertoires than other groups. Despite higher TCR clonal expansions in HAM/TSP patients, no disease-specific TCRs were shared among patients. Only non-shared or "private" TCR repertoires was observed. While no clones that shared the same CDR3 amino acid sequences were seen in either HC or MS patients, there was a cluster of related CDR3 amino acid sequences observed for 18 out of 34 MS patients when evaluated by phylogenetic tree analysis. This suggests that a TCR-repertoire signature may be identified in a subset of patients with MS.
在这项研究中，我们对包括人类 T 细胞嗜淋巴细胞病毒 I 型（HTLV-I）感染相关的慢性进行性炎症性神经疾病（HAM/TSP）和多发性硬化症（MS）在内的慢性进行性炎症性神经疾病患者的 TCR 受体谱特征进行了研究。MS 是一种病因不明的中枢神经系统炎症性脱髓鞘疾病。我们假设 T 细胞受体（TCR）克隆谱“特征”可以将 HAM/TSP 患者与健康对照者以及与 MS 等更具异质性中枢神经系统反应性炎症性疾病的患者区分开来。在这项研究中，我们应用了一种无偏分子技术 - 基于独特分子标识符（UMI）文库的策略，通过高通量测序（HTS）技术以高精度研究 TCR 克隆谱。从 14 名 HAM/TSP 患者、34 名 MS 患者和 20 名健康对照者（HC）的 200 万个外周单核细胞（PBMC）中测序了 cDNA-TCR β 链文库。与 MS 和 HC 相比，HAM/TSP 患者的克隆 T 细胞扩增更高，而 TCR 克隆扩增的增加与所有受试者 TCR 谱的多样性呈反比。此外，对 HAM/TSP 患者 TCR 谱的纵向分析表明，TCR 克隆扩增与 HTLV-I 前病毒载量相关。令人惊讶的是，MS 患者的 TCR 谱多样性高于其他组。尽管 HAM/TSP 患者的 TCR 克隆扩增更高，但患者之间没有共享的疾病特异性 TCR。只观察到非共享或“私有”TCR 谱。虽然在 HC 或 MS 患者中未观察到共享相同 CDR3 氨基酸序列的克隆，但通过系统发育树分析，在 34 名 MS 患者中有 18 名观察到相关的 CDR3 氨基酸序列簇。这表明在 MS 的一部分患者中可能确定 TCR 受体谱特征。
