Insertion of the Dibasic Motif in the Flanking Region of a Cryptic Self-Determinant Leads to Activation of the Epitope-Specific T Cells

• Published in: The Journal of immunology (1950) 2005-08, Vol.175 (4), p.2252-2260
• Main Authors: Zhu, Hui, Liu, Kechang, Cerny, Jan, Imoto, Taiji, Moudgil, Kamal D
• Format: Article
• Language: English
• Subjects: Amino Acid Motifs - genetics; Amino Acid Sequence; Animals; Antigen Presentation - genetics; Aspartic Acid - genetics; Autoantibodies - biosynthesis; Autoantibodies - metabolism; Autoantigens - administration & dosage; Autoantigens - genetics; Autoantigens - immunology; Autoantigens - metabolism; Cross Reactions - genetics; Female; H-2 Antigens - immunology; Immunodominant Epitopes - administration & dosage; Immunodominant Epitopes - genetics; Immunodominant Epitopes - immunology; Immunodominant Epitopes - metabolism; Lymphocyte Activation - genetics; Lysine - genetics; Mice; Mice, Inbred BALB C; Mice, Inbred C3H; Mice, Inbred C57BL; Molecular Sequence Data; Muramidase - administration & dosage; Muramidase - genetics; Muramidase - immunology; Muramidase - metabolism; Mutagenesis, Site-Directed; Peptide Fragments - administration & dosage; Peptide Fragments - genetics; Peptide Fragments - immunology; Peptide Fragments - metabolism; Self Tolerance - genetics
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.175.4.2252

Efficient induction of self tolerance is critical for avoiding autoimmunity. The T cells specific for the well-processed and -presented (dominant) determinants of a native self protein are generally tolerized in the thymus, whereas those potentially directed against the inefficiently processed and presented (cryptic) self epitopes escape tolerance induction. We examined whether the crypticity of certain determinants of mouse lysozyme-M (ML-M) could be attributed to the nonavailability of a proteolytic site, and whether it could be reversed to immunodominance by engraftment of a novel cleavage site in the flanking region of the epitope. Using site-directed mutagenesis, we created the dibasic motif (RR or RK; R = arginine, K = lysine), a target of intracellular proteases, in the region adjoining one of the three cryptic epitopes (46-61, 66-79, or 105-119) of ML-M. Interestingly, the mutated lysozyme proteins, but not unmutated ML-M, were immunogenic in mice. The T cell response to the altered lysozyme was attributable to the efficient processing and presentation of the previously cryptic epitope, and this response was both epitope and MHC haplotype specific. In addition, the anti-self T cell response was associated with the generation of autoantibodies against self lysozyme. However, the results using one of three mutated lysozymes suggested that the naturally processed, dibasic motif-marked epitope may not always correspond precisely to the cryptic determinant within a synthetic peptide. This is the first report describing the circumvention of self tolerance owing to the targeted reversal of crypticity to dominance in vivo of a specific epitope within a native self Ag.