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Cell Rep. 2017 Aug 1;20(5):1242-1253. doi: 10.1016/j.celrep.2017.07.026.

Multi-level Strategy for Identifying Proteasome-Catalyzed Spliced Epitopes Targeted by CD8+ T Cells during Bacterial Infection.

Author information

1
Division of Immunology, Faculty of Veterinary Medicine, Utrecht University, 3571 EK Utrecht, the Netherlands.
2
Centre for Integrative Systems Biology and Bioinformatics, Department of Life Sciences, Imperial College London, SW7 2AZ London, UK; Max-Planck-Institute for Biophysical Chemistry, 37077 Göttingen, Germany.
3
Institut für Biochemie, Charité - Universitätsmedizin Berlin, 10117 Berlin, Germany; Berlin Institute of Health, 10117 Berlin, Germany; Shared Facility for Mass Spectrometry, Charité - Universitätsmedizin Berlin, 10117 Berlin, Germany.
4
Institut für Biochemie, Charité - Universitätsmedizin Berlin, 10117 Berlin, Germany; Berlin Institute of Health, 10117 Berlin, Germany.
5
Institut für Biochemie, Charité - Universitätsmedizin Berlin, 10117 Berlin, Germany.
6
Institut für Biochemie, Charité - Universitätsmedizin Berlin, 10117 Berlin, Germany; Berlin Institute of Health, 10117 Berlin, Germany; Centre for Inflammation Biology and Cancer Immunology (CIBCI) & Peter Gorer Department of Immunobiology, King's College London, SE1 1UL London, UK. Electronic address: michele.mishto@kcl.ac.uk.
7
Division of Immunology, Faculty of Veterinary Medicine, Utrecht University, 3571 EK Utrecht, the Netherlands. Electronic address: e.j.a.m.sijts@uu.nl.

Abstract

Proteasome-catalyzed peptide splicing (PCPS) generates peptides that are presented by MHC class I molecules, but because their identification is challenging, the immunological relevance of spliced peptides remains unclear. Here, we developed a reverse immunology-based multi-level approach to identify proteasome-generated spliced epitopes. Applying this strategy to a murine Listeria monocytogenes infection model, we identified two spliced epitopes within the secreted bacterial phospholipase PlcB that primed antigen-specific CD8+ T cells in L. monocytogenes-infected mice. While reacting to the spliced epitopes, these CD8+ T cells failed to recognize the non-spliced peptide parts in the context of their natural flanking sequences. Thus, we here show that PCPS expands the CD8+ T cell response against L. monocytogenes by exposing spliced epitopes on the cell surface. Moreover, our multi-level strategy opens up opportunities to systematically investigate proteins for spliced epitope candidates and thus strategies for immunotherapies or vaccine design.

KEYWORDS:

Listeria monocytogenes; antigen presentation; in silico analysis; intracelllular bacteria; peptide splicing; proteasome

PMID:
28768206
DOI:
10.1016/j.celrep.2017.07.026
[Indexed for MEDLINE]
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