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While most of such epitopes are generated by proteasome\u2010mediated antigen cleavage, analysis of tumor antigen processing has revealed that epitopes may also derive from proteasome\u2010catalyzed peptide splicing (PCPS). To determine whether PCPS contributes to epitope processing during infection, we analyzed the fragments produced by purified proteasomes from a <jats:italic>Listeria monocytogenes<\/jats:italic> polypeptide. Mass spectrometry identified a known H\u20102K<jats:sup>b<\/jats:sup>\u2010presented linear epitope (LLO<jats:sub>296\u2010304<\/jats:sub>) in the digests, as well as four spliced peptides that were trimmed by ERAP into peptides with in silico predicted H\u20102K<jats:sup>b<\/jats:sup> binding affinity. These spliced peptides, which displayed sequence similarity with LLO<jats:sub>296\u2010304<\/jats:sub>, bound to H\u20102K<jats:sup>b<\/jats:sup> molecules in cellular assays and one of the peptides was recognized by CD8<jats:sup>+<\/jats:sup> T\u00a0cells of infected mice. This spliced epitope differed by one amino acid from LLO<jats:sub>296\u2010304<\/jats:sub> and double staining with LLO<jats:sub>296\u2010304<\/jats:sub>\u2010 and spliced peptide\u2010folded MHC multimers showed that LLO<jats:sub>296\u2010304<\/jats:sub> and its spliced variant were recognized by the same CD8<jats:sup>+<\/jats:sup> T\u00a0cells. Thus, PCPS multiplies the variety of peptides that is processed from an antigen and leads to the production of epitope variants that can be recognized by cross\u2010reacting pathogen\u2010specific CD8<jats:sup>+<\/jats:sup> T\u00a0cells. Such mechanism may reduce the chances for pathogen immune evasion.<\/jats:p>","DOI":"10.1002\/eji.201545989","type":"journal-article","created":{"date-parts":[[2016,2,25]],"date-time":"2016-02-25T04:41:54Z","timestamp":1456375314000},"page":"1109-1118","source":"Crossref","is-referenced-by-count":43,"title":["CD8<sup>+<\/sup> T\u00a0cells of <i>Listeria monocytogenes\u2010<\/i>infected mice recognize both linear and spliced proteasome products"],"prefix":"10.1002","volume":"46","author":[{"given":"Anouk C. 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