title: Absence of recognition of common melanocytic antigens by T cells isolated from the cerebrospinal fluid of a Vogt-Koyanagi-Harada patient.
abstract: OBJECTIVE
  Vogt-Koyanagi-Harada (VKH) syndrome is an autoimmune disease characterized by inaugural uveomeningitidis and hearing loss and at late stages a depigmentation in eyes and skin. Melanocytes are the cells common to the four affected tissues, namely eye, brain, inner ear, and skin. Melanocytes are therefore considered as the source of self-antigens. The melanocytic proteins tyrosinase-related protein-1 (TRP1), TRP2, tyrosinase, and gp100 have been proposed as the proteins targeted by autoreactive T cells from VKH patients bearing human leukocyte antigen (HLA)-DRB1*04:05, the HLA allele classically associated with VKH disease. The objective of this work was to determine the antigens recognized by a large number of potentially autoreactive CD4 T lymphocytes obtained from the cerebrospinal fluid of one VKH patient who did not express HLA-DRB1*04:05.
  
  
  METHODS
  T cells were isolated from the cerebrospinal fluid of a newly diagnosed HLA-DRB1*14:01,*15:03;-DPB1*01:01,*04:02 patient in the acute phase of the VKH disease and cloned by limiting dilution. Each of the 107 T cell clones, of which 90% were CD4(+), was tested for its ability to secrete cytokines upon contact with autologous antigen-presenting cells loaded with either of the melanocytic proteins TRP1, TRP2, tyrosinase, gp100, Melan-A and KU-MEL-1. The sensitivity of our recombinant bacteria-based approach was validated with a CD4 T cell clone with known antigen specificity. The ability of each of the 107 clones to secrete cytokines upon nonspecific stimulation was verified.
  
  
  RESULTS
  None of the 107 T cell clones was able to secrete tumor necrosis factor-α, interferon-γ, interleukin (IL)-5, or IL-17 upon contact with autologous B cells loaded with any of the six common melanocytic proteins. Nine clones secreted high-level IL-17 upon stimulation with beads coated with antibodies.
  
  
  CONCLUSIONS
  The self-antigens that triggered the VKH disease in this patient probably derive from proteins other than the six melanocytic proteins mentioned above. Further study of antigens that are recognized by potential autoreactive T cells from VKH patients is likely to benefit from testing a broader set of melanocytic proteins.
authors.affiliation: Université Paris 13, Sorbonne Paris Cité, Laboratoire de recherche clinique et thérapeutique, Bobigny, France; Service de Médecine Interne, Hôpital Avicenne, Assistance Publique-Hôpitaux de Paris (AP-HP), Bobigny, France.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Abad
authors.affiliation: Ludwig Institute for Cancer Research and de Duve Institute, Université catholique de Louvain, 74 av. Hippocrate, Brussels, Belgium.
authors.forename: Grégoire
authors.initials: G
authors.lastname: Wieërs
authors.affiliation: Ludwig Institute for Cancer Research and de Duve Institute, Université catholique de Louvain, 74 av. Hippocrate, Brussels, Belgium.
authors.forename: Didier
authors.initials: D
authors.lastname: Colau
authors.affiliation: Ludwig Institute for Cancer Research and de Duve Institute, Université catholique de Louvain, 74 av. Hippocrate, Brussels, Belgium.
authors.forename: Claude
authors.initials: C
authors.lastname: Wildmann
authors.affiliation: Université Paris Descartes, Faculté de Médecine, AP-HP, Groupe Hospitalier Cochin-Hôtel-Dieu, Paris, France.
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Delair
authors.affiliation: Université Paris 13, Sorbonne Paris Cité, Laboratoire de recherche clinique et thérapeutique, Bobigny, France; Service de Médecine Interne, Hôpital Avicenne, Assistance Publique-Hôpitaux de Paris (AP-HP), Bobigny, France.
authors.forename: Robin
authors.initials: R
authors.lastname: Dhote
authors.affiliation: Université Paris Descartes, Faculté de Médecine, AP-HP, Groupe Hospitalier Cochin-Hôtel-Dieu, Paris, France.
authors.forename: Antoine P
authors.initials: AP
authors.lastname: Brézin
authors.affiliation: Division of Cellular Signaling, Institute for Advanced Medical Research, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan.
authors.forename: Yukata
authors.initials: Y
authors.lastname: Kawakami
authors.affiliation: de Duve Institute, Université catholique de Louvain, Brussels, Belgium.
authors.forename: Pierre G
authors.initials: PG
authors.lastname: Coulie
authors.affiliation: Ludwig Institute for Cancer Research and de Duve Institute, Université catholique de Louvain, 74 av. Hippocrate, Brussels, Belgium.
authors.forename: Pierre
authors.initials: P
authors.lastname: van der Bruggen
mesh_terms: D000328:Adult; D000951:Antigens, Neoplasm / Q000378:metabolism; D001324:Autoantigens / Q000276:immunology*; D001402:B-Lymphocytes / Q000821:virology; D001419:Bacteria / Q000378:metabolism; D015153:Blotting, Western; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D002469:Cell Separation; D002999:Clone Cells; D016207:Cytokines / Q000378:metabolism; D000939:Epitopes / Q000276:immunology; D004854:Herpesvirus 4, Human / Q000276:immunology; D006801:Humans; D020381:Interleukin-17 / Q000378:metabolism; D008297:Male; D008544:Melanocytes / Q000276:immunology*; D009363:Neoplasm Proteins / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology*; D014607:Uveomeningoencephalitic Syndrome / Q000134:cerebrospinal fluid* / Q000276:immunology* / Q000473:pathology
chemical_list: D000951:Antigens, Neoplasm; D001324:Autoantigens; D016207:Cytokines; D000939:Epitopes; D020381:Interleukin-17; C072013:MAGEA3 protein, human; D009363:Neoplasm Proteins; D011994:Recombinant Proteins