title: High-throughput sequencing of TCR repertoires in multiple sclerosis reveals intrathecal enrichment of EBV-reactive CD8+ T cells.
abstract: 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.
authors.affiliation: 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.
authors.forename: Andreas
authors.initials: A
authors.lastname: Lossius
authors.forename: Jorunn N
authors.initials: JN
authors.lastname: Johansen
authors.forename: Frode
authors.initials: F
authors.lastname: Vartdal
authors.forename: Harlan
authors.initials: H
authors.lastname: Robins
authors.forename: Benth
authors.initials: B
authors.lastname: Jūratė Šaltytė
authors.forename: Trygve
authors.initials: T
authors.lastname: Holmøy
authors.forename: Johanna
authors.initials: J
authors.lastname: Olweus
mesh_terms: D000328:Adult; D001402:B-Lymphocytes / Q000276:immunology / Q000821:virology; D001483:Base Sequence; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology* / Q000821:virology; D005260:Female; D015234:HLA-A Antigens / Q000276:immunology; D015235:HLA-B Antigens / Q000276:immunology; D059811:HLA-DRB1 Chains / Q000276:immunology; D004854:Herpesvirus 4, Human / Q000276:immunology*; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D007244:Infectious Mononucleosis / Q000276:immunology / Q000821:virology; D008297:Male; D008875:Middle Aged; D009103:Multiple Sclerosis / Q000097:blood / Q000134:cerebrospinal fluid / Q000276:immunology* / Q000821:virology; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000097:blood* / Q000235:genetics / Q000276:immunology; D017422:Sequence Analysis, DNA; D055815:Young Adult
chemical_list: D015234:HLA-A Antigens; D015235:HLA-B Antigens; D059811:HLA-DRB1 Chains; D016693:Receptors, Antigen, T-Cell, alpha-beta
keywords: Cerebrospinal fluid; Epstein-Barr virus; High-throughput sequencing; MS; TCR