It has been argued that tumor progenitor cells play a crucial role in tumor development

l as the I-Ak-restricted hen egg-white lysozyme epitopes. Both studies demonstrated unequivocally that conversion of tyrosine to nitrotyrosine resulted in dramatic consequences for T cell recognition. Indeed, processing of native proteins by activated antigen presenting cells resulted in the presentation of MHC class II-restricted NT-epitopes in lymphoid tissues, AG1024 biological activity significantly altering specific T cell responses and eliciting NT-specific CD4+ T cells that evaded negative selection in the thymus and thus central tolerance. Likewise MHC-I-Restricted Nitrotyrosinated Neoantigen loss of tolerance by NT-specific CD4+ T cells has recently been shown to be critical for the production of autoreactive antibodies. These studies indicate that NT-proteins generated during inflammation might constitute an important class of neoantigens that could promote autoimmune T cell responses. The well-established lymphocytic choriomeningitis virus glycoprotein system was also used by Hardy et al to demonstrate that conversion of tyrosine to NY also profoundly affected T cell recognition of MHC class I-restricted epitopes. A significant amount of the overall CD8+ T cell response to LCMV is dominated by very few viral epitopes, comprising the H2Db-and H-2Kb- restricted peptide gp3341 and the H-2Kb-restricted peptide gp34 41 . Both gp33 and gp34 contain a single tyrosine residue at positions 4 and 3, respectively. T cell populations, exclusively specific to the nitrotyrosinated MHC complexes H-2Db/NY-gp33, H-2Kb/NYgp33 and/or H-2Kb/NY-gp34, were elicited in C57/BL6 mice following immunization with the nitrated peptide NY-gp33. Importantly, CD8+ T cell hybridomas specific for NY-gp33 comprised two distinct subsets recognizing either H2Db/NY-gp33 or H-2Kb/NY-gp33. While the T cell hybridoma 24H1 responded to stimulation with both H-2Kb/NY-gp34 andH2Kb/NY-gp33, it did not recognize the unmodified wild-type MHC complex H-2Kb/gp34 nor H-2Kb/gp33. Similarly, the H-2Db/NY-gp33-specific T cell hybridoma 4C8 did not recognize H-2Db/gp33. In contrast, nitrotyrosination of the main T cell receptor -interacting peptide tyrosine residue p4Y abrogated recognition of H-2Db/NY-gp33 MHC complexes by H-2Db/ gp33-specific T cells from P14 transgenic mice. The present study provides a comparative biochemical and structural analysis that explains how nitrotyrosination of the LCMV-associated immunodominant epitopes gp33 and gp34 can alter T cell recognition in the context of the two different MHC class I molecules H-2Db and H-2Kb. Nitrotyrosination of the MHC-restricted peptide impairs TCR recognition through reduced stability and alteration of the molecular surface of the MHC complex. The possible implications for the role of nitrotyrosination in the creation of modified neoantigens that allow for viral escape and/or breaking of immune tolerance that possibly results in autoimmune disorders are discussed. 180 and 360 images were collected with 0.5u oscillation per frame for both H-2Kb/gp34 and H-2Kb/NY-gp34. Data were processed with MOSFLM and SCALA from the CCP4 suite. The space group was determined to be P212121 PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/22189254 for H-2Kb/gp34 with unit-cell parameters a = 88.4, b = 92.6, c = 128.8 A. The space b group for H-2K /NY-gp34 wasP21 with unit-cell parameters a = 50.5, b = 88.5, c = 119 A, a = c = 90.0u and b = 94.7u. Data collection statistics are provided in Crystal structure determination and refinement The crystal structure of H-2Kb/gp34 was solved by molecular replacement using Phase