PMID- 24991188 OWN - NLM STAT- MEDLINE DCOM- 20140930 LR - 20240321 IS - 1090-0535 (Electronic) IS - 1090-0535 (Linking) VI - 20 DP - 2014 TI - Absence of recognition of common melanocytic antigens by T cells isolated from the cerebrospinal fluid of a Vogt-Koyanagi-Harada patient. PG - 956-69 AB - PURPOSE: 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-alpha, interferon-gamma, 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. FAU - Abad, Sebastien AU - Abad S AD - Universite Paris 13, Sorbonne Paris Cite, Laboratoire de recherche clinique et therapeutique, Bobigny, France; Service de Medecine Interne, Hopital Avicenne, Assistance Publique-Hopitaux de Paris (AP-HP), Bobigny, France. FAU - Wieers, Gregoire AU - Wieers G AD - Ludwig Institute for Cancer Research and de Duve Institute, Universite catholique de Louvain, 74 av. Hippocrate, Brussels, Belgium. FAU - Colau, Didier AU - Colau D AD - Ludwig Institute for Cancer Research and de Duve Institute, Universite catholique de Louvain, 74 av. Hippocrate, Brussels, Belgium. FAU - Wildmann, Claude AU - Wildmann C AD - Ludwig Institute for Cancer Research and de Duve Institute, Universite catholique de Louvain, 74 av. Hippocrate, Brussels, Belgium. FAU - Delair, Emmanuelle AU - Delair E AD - Universite Paris Descartes, Faculte de Medecine, AP-HP, Groupe Hospitalier Cochin-Hotel-Dieu, Paris, France. FAU - Dhote, Robin AU - Dhote R AD - Universite Paris 13, Sorbonne Paris Cite, Laboratoire de recherche clinique et therapeutique, Bobigny, France; Service de Medecine Interne, Hopital Avicenne, Assistance Publique-Hopitaux de Paris (AP-HP), Bobigny, France. FAU - Brezin, Antoine P AU - Brezin AP AD - Universite Paris Descartes, Faculte de Medecine, AP-HP, Groupe Hospitalier Cochin-Hotel-Dieu, Paris, France. FAU - Kawakami, Yukata AU - Kawakami Y AD - Division of Cellular Signaling, Institute for Advanced Medical Research, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan. FAU - Coulie, Pierre G AU - Coulie PG AD - de Duve Institute, Universite catholique de Louvain, Brussels, Belgium. FAU - van der Bruggen, Pierre AU - van der Bruggen P AD - Ludwig Institute for Cancer Research and de Duve Institute, Universite catholique de Louvain, 74 av. Hippocrate, Brussels, Belgium. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20140702 PL - United States TA - Mol Vis JT - Molecular vision JID - 9605351 RN - 0 (Antigens, Neoplasm) RN - 0 (Autoantigens) RN - 0 (Cytokines) RN - 0 (Epitopes) RN - 0 (Interleukin-17) RN - 0 (MAGEA3 protein, human) RN - 0 (Neoplasm Proteins) RN - 0 (Recombinant Proteins) SB - IM MH - Adult MH - Antigens, Neoplasm/metabolism MH - Autoantigens/*immunology MH - B-Lymphocytes/virology MH - Bacteria/metabolism MH - Blotting, Western MH - CD4-Positive T-Lymphocytes/immunology MH - Cell Separation MH - Clone Cells MH - Cytokines/metabolism MH - Epitopes/immunology MH - Herpesvirus 4, Human/immunology MH - Humans MH - Interleukin-17/metabolism MH - Male MH - Melanocytes/*immunology MH - Neoplasm Proteins/metabolism MH - Recombinant Proteins/metabolism MH - T-Lymphocytes/*immunology MH - Uveomeningoencephalitic Syndrome/*cerebrospinal fluid/*immunology/pathology PMC - PMC4077848 EDAT- 2014/07/06 06:00 MHDA- 2014/10/01 06:00 PMCR- 2014/01/01 CRDT- 2014/07/04 06:00 PHST- 2014/04/03 00:00 [received] PHST- 2014/06/30 00:00 [accepted] PHST- 2014/07/04 06:00 [entrez] PHST- 2014/07/06 06:00 [pubmed] PHST- 2014/10/01 06:00 [medline] PHST- 2014/01/01 00:00 [pmc-release] AID - 91 [pii] PST - epublish SO - Mol Vis. 2014 Jul 2;20:956-69. eCollection 2014.