title: Identification of an HLA-A*0201-restricted epitopic peptide from human dystrophin: application in duchenne muscular dystrophy gene therapy.
abstract: Dystrophin-based gene therapy treatments aimed at correcting the Duchenne muscular dystrophy phenotype require stable expression of normal dystrophin (DYST) protein in myocytes without immune responses, which would compromise long-term expression. To predict cytotoxic T-cell-mediated responses elicited by transgenes, we used here H-2-negative HLA-A*0201 transgenic mice and identified human DYST epitopes, which elicit HLA-A*0201-restricted cytotoxic T cell activities. Among a series of eight peptides predicted from the human DYST sequence, not shared with the endogenous mouse DYST sequence, four of them were able to bind to HLA-A*0201 molecules and to induce cytotoxic T lymphocyte (CTL) responses. After human DYST DNA transfer in muscle of HLA-A*0201 mice, only the human DYST1281 epitope, located in the spectrin-like repeat 9 domain, induced strong CD8(+) CTL responses. Using the corresponding human DYST1281 peptide/HLA-A*0201 tetramer, we detected human DYST1281-specific CD8(+) T cells in peripheral lymphoid organs and blood of HLA-A*0201 mice injected with human DYST DNA. Our results demonstrate that muscle injection with human DYST DNA systematically triggers CTL responses against this HLA-A*0201-restricted human DYST1281 peptide, which is present in long human DYST isoforms. Identification of such immunodominant human DYST epitopes and use of peptide/HLA tetramers will allow the immunomonitoring of CTL responses in HLA-phenotyped Duchenne muscular dystrophy patients undergoing gene therapy. Finally, the knowledge of HLA-A*0201-restricted human DYST peptides will be of importance to test, in mouse models, new immunomodulatory interventions allowing long-term engraftment of human dystrophin.
authors.affiliation: Laboratoire d'Immunologie, Généthon, CNRS UMR 8115, 91002, Evry Cedex, France. davoust@genethon.fr
authors.forename: F
authors.initials: F
authors.lastname: Ginhoux
authors.forename: C
authors.initials: C
authors.lastname: Doucet
authors.forename: M
authors.initials: M
authors.lastname: Leboeuf
authors.forename: F A
authors.initials: FA
authors.lastname: Lemonnier
authors.forename: O
authors.initials: O
authors.lastname: Danos
authors.forename: J
authors.initials: J
authors.lastname: Davoust
authors.forename: H
authors.initials: H
authors.lastname: Firat
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D016189:Dystrophin / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D000939:Epitopes / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D005434:Flow Cytometry; D015316:Genetic Therapy / Q000379:methods*; D015234:HLA-A Antigens / Q000276:immunology*; D015789:HLA-A2 Antigen; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics* / Q000276:immunology / Q000628:therapy*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
chemical_list: D016189:Dystrophin; D000939:Epitopes; D015234:HLA-A Antigens; C435939:HLA-A*02:01 antigen; D015789:HLA-A2 Antigen