title: Tolerance induction by intrathymic expression of P0.
abstract: Genetic deficiency or instability of myelin protein zero (P0) results in hereditary motor sensory neuropathy. In view of recent advances in gene therapy, substitution of the molecular defect may become realistic in the near future. Here we investigate the impact of genetic deficiency of P0 on selection of the autoreactive T cell repertoire in the corresponding mouse model. We show that P0 mRNA transcripts are expressed in thymic stroma, similar to other myelin proteins and that expression of intact P0 protein can be detected by Western blot. Using a library of overlapping 20mer peptides spanning the entire length of P0 and applying the ELISPOT technique, we detected a strong immune response toward P0 extracellular domain peptide aa 41-60 in P0(-/-) knockout mice, but not in heterozygous P0(+/-) or wild-type (wt) mice. In addition, one cryptic epitope and two subdominant epitopes of P0 were identified. Using P0(-/-) into wt bone marrow (BM) chimeras we found that P0 expression in the host suffices for full tolerance induction, which is in line with its presence in thymic stroma. However, repopulation of P0(-/-) mice with wt BM led to partial induction of tolerance, suggesting that BM derived cells can also express this protein. Our findings may have implications for secondary autoimmunity developing after gene therapy in hereditary neuropathies and other diseases with genetically determined protein deficiency, because the repaired protein will then represent a foreign, nontolerized Ag.
authors.affiliation: Department of Neurology, Clinical Research Group for Multiple Sclerosis and Neuroimmunology, University of Würzburg, Würzburg, Germany.
authors.forename: Lucian
authors.initials: L
authors.lastname: Visan
authors.forename: Ioana A
authors.initials: IA
authors.lastname: Visan
authors.forename: Andreas
authors.initials: A
authors.lastname: Weishaupt
authors.forename: Harald H
authors.initials: HH
authors.lastname: Hofstetter
authors.forename: Klaus V
authors.initials: KV
authors.lastname: Toyka
authors.forename: Thomas
authors.initials: T
authors.lastname: Hünig
authors.forename: Ralf
authors.initials: R
authors.lastname: Gold
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017951:Antigen Presentation / Q000235:genetics; D001854:Bone Marrow Cells / Q000276:immunology / Q000378:metabolism / Q000528:radiation effects; D002454:Cell Differentiation / Q000235:genetics / Q000276:immunology; D018984:Epitopes, T-Lymphocyte / Q000008:administration & dosage / Q000032:analysis / Q000378:metabolism; D005333:Fetus; D018628:Gene Dosage; D006412:Hematopoietic Stem Cells / Q000276:immunology / Q000378:metabolism / Q000528:radiation effects; D007108:Immune Tolerance / Q000235:genetics; D016056:Immunodominant Epitopes / Q000008:administration & dosage / Q000032:analysis / Q000378:metabolism; D007156:Immunologic Memory / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008969:Molecular Sequence Data; D018993:Myelin P0 Protein / Q000096:biosynthesis* / Q000172:deficiency / Q000276:immunology / Q000378:metabolism; D009924:Organ Culture Techniques; D010446:Peptide Fragments / Q000008:administration & dosage / Q000276:immunology / Q000378:metabolism; D011828:Radiation Chimera / Q000276:immunology; D016176:T-Lymphocyte Subsets / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D013950:Thymus Gland / Q000166:cytology / Q000276:immunology* / Q000378:metabolism*
chemical_list: D018984:Epitopes, T-Lymphocyte; D016056:Immunodominant Epitopes; D018993:Myelin P0 Protein; D010446:Peptide Fragments