Quantifying epitope presentation using mass spectrometry

Croft, Nathan P.; Purcell, Anthony W.; Tscharke, David C.

doi:10.1016/j.molimm.2015.06.010

Cited by 33 publications

(28 citation statements)

References 25 publications

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“…Such tight linkage between viral protein synthesis and peptide generation is the rule and not the exception, as shown by a mounting number of studies (reviewed in Anton and Yewdell (4)). This is particularly well shown by applying mass spectrometry to kinetic measurements of viral protein vs. peptide generation, which enables the simultaneous characterization of dozens of gene products (5, 40), revealing the dominant contribution of DRiPs to peptide generation from viral gene products.…”

Section: Discussionmentioning

confidence: 98%

Defining Viral Defective Ribosomal Products: Standard and Alternative Translation Initiation Events Generate a Common Peptide from Influenza A Virus M2 and M1 mRNAs

Yang

Gibbs

Hickman

et al. 2016

The Journal of Immunology

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Influenza A virus gene segment 7 encodes two proteins: the M1 protein translated from unspliced mRNA and the M2 protein produced by mRNA splicing and largely encoded by the M1 +1 reading frame. To better understand the generation of defective ribosomal products (DRiPs) relevant to MHC class I antigen presentation, we engineered influenza A virus gene segment 7 to encode the model H-2 Kb class I peptide ligand SIINFEKL at the M2 protein C-terminus. Remarkably, after treating virus-infected cells with the RNA splicing inhibitor spliceostatin A to prevent M2 mRNA generation, Kb-SIINFEKL complexes were still presented on the cell surface at levels of up to 60% of untreated cells. Three key findings indicate that SIINFEKL is produced by cytoplasmic translation of unspliced M1 mRNA initiating at CUG codons within the +1 reading frame: Synonymous mutation of CUG codons in the M2-reading frame reduced Kb-SIINFEKL generation.Kb-SIINFEKL generation was not affected by drug-mediated inhibition of AUG-initiated M1 synthesis.Kb-SIINFEKL was generated in vitro and in vivo from mRNA synthesized in the cytoplasm by vaccinia virus, and hence cannot be spliced. These findings define a viral DRiP generated by cytoplasmic non-canonical translation and demonstrate the participation of CUG-codon-based translation initiation in pathogen immunosurveillance.

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Section: Discussionmentioning

confidence: 98%

Defining Viral Defective Ribosomal Products: Standard and Alternative Translation Initiation Events Generate a Common Peptide from Influenza A Virus M2 and M1 mRNAs

Yang

Gibbs

Hickman

et al. 2016

The Journal of Immunology

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“…Because of the increased role of DM in MHC II processing under inflammatory conditions, the abundance of many self-peptides displayed at the cell surface is likely to change, with implications for the regulation of self-tolerance. Methodology for the reliable quantitative measurement of peptide abundances in complex samples is available, and very recently has begun to be applied to immunopeptidomes [105,106]. Thus we hope to learn in even greater detail about the distribution of peptides in the MHC II melting pot, and how they regulate T cell activity.…”

Section: Discussionmentioning

confidence: 99%

The melting pot of the MHC II peptidome

Stern

Santambrogio

2016

Current Opinion in Immunology

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Recent advances in mass spectrometry technology have facilitated detailed examination of MHC-II immunopeptidomes, i.e. the repertoires of peptides bound to MHC-II molecules expressed in antigen presenting cells. These studies have deepened our view of MHC-II presentation. Other studies have broadened our view of pathways leading up to peptide loading. Here we review these recent studies in the context of earlier work on conventional and non-conventional MHC-II processing. The message that emerges is that sources of antigen beyond conventional endosomal processing of endocytosed proteins are important for generation of cellular immune responses to pathogens and maintenance of central and peripheral tolerance. The multiplicity of pathways results in a broad MHC II immunopeptidome that conveys the sampled environment to patrolling T cells.

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“…Rapid advances in mass spectrometry instrumentation and methods are enabling both the relative and absolute quantification of pMHC complexes on in vitro cells, and are bringing the capacity to accurately quantify 72 . Whereas traditional data-dependent mass spectrometry methods provide ideal workflows for epitope discovery, the gold standard in quantification is the ability to target specific peptide sequences for detection through a process termed multiple reaction monitoring (MRM).…”

Section: Interactions Between T Cells Of Different Specificitiesmentioning

confidence: 99%

Sizing up the key determinants of the CD8+ T cell response

et al. 2015

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Naive CD8(+) T cells give rise to cytotoxic T lymphocytes (CTLs), which promote the effective eradication of viruses and tumours. Although the past decades have seen enormous advances in cellular immunology, a precise understanding of the key elements that determine the specificity and magnitude of primary CTL responses has been lacking. However, recent technological advances have allowed us to more accurately identify, characterize and quantitate key determinants that define the specificity and magnitude of CD8(+) T cell-mediated immunity. This Review discusses the technical and conceptual advances that have markedly changed our understanding of the determinants of primary CTL responses.

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Quantifying epitope presentation using mass spectrometry

Cited by 33 publications

References 25 publications

Defining Viral Defective Ribosomal Products: Standard and Alternative Translation Initiation Events Generate a Common Peptide from Influenza A Virus M2 and M1 mRNAs

Defining Viral Defective Ribosomal Products: Standard and Alternative Translation Initiation Events Generate a Common Peptide from Influenza A Virus M2 and M1 mRNAs

The melting pot of the MHC II peptidome

Sizing up the key determinants of the CD8+ T cell response

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