Mass spectrometry of human leukocyte antigen class I peptidomes reveals strong effects of protein abundance and turnover on antigen presentation.

Details

Serval ID
serval:BIB_69D719AEE7F3
Type
Article: article from journal or magazin.
Collection
Publications
Title
Mass spectrometry of human leukocyte antigen class I peptidomes reveals strong effects of protein abundance and turnover on antigen presentation.
Journal
Molecular & cellular proteomics
Author(s)
Bassani-Sternberg M., Pletscher-Frankild S., Jensen L.J., Mann M.
ISSN
1535-9484 (Electronic)
ISSN-L
1535-9476
Publication state
Published
Issued date
03/2015
Peer-reviewed
Oui
Volume
14
Number
3
Pages
658-673
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Abstract
HLA class I molecules reflect the health state of cells to cytotoxic T cells by presenting a repertoire of endogenously derived peptides. However, the extent to which the proteome shapes the peptidome is still largely unknown. Here we present a high-throughput mass-spectrometry-based workflow that allows stringent and accurate identification of thousands of such peptides and direct determination of binding motifs. Applying the workflow to seven cancer cell lines and primary cells, yielded more than 22,000 unique HLA peptides across different allelic binding specificities. By computing a score representing the HLA-I sampling density, we show a strong link between protein abundance and HLA-presentation (p < 0.0001). When analyzing overpresented proteins - those with at least fivefold higher density score than expected for their abundance - we noticed that they are degraded almost 3 h faster than similar but nonpresented proteins (top 20% abundance class; median half-life 20.8h versus 23.6h, p < 0.0001). This validates protein degradation as an important factor for HLA presentation. Ribosomal, mitochondrial respiratory chain, and nucleosomal proteins are particularly well presented. Taking a set of proteins associated with cancer, we compared the predicted immunogenicity of previously validated T-cell epitopes with other peptides from these proteins in our data set. The validated epitopes indeed tend to have higher immunogenic scores than the other detected HLA peptides. Remarkably, we identified five mutated peptides from a human colon cancer cell line, which have very recently been predicted to be HLA-I binders. Altogether, we demonstrate the usefulness of combining MS-analysis with immunogenesis prediction for identifying, ranking, and selecting peptides for therapeutic use.
Keywords
Antigen Presentation, Cell Line, Tumor, Cells, Cultured, Epitopes, T-Lymphocyte/metabolism, HCT116 Cells, HLA Antigens/metabolism, Histocompatibility Antigens Class I/immunology, Humans, Mass Spectrometry/methods, Neoplasms/immunology, Peptides/immunology, Peptides/isolation & purification, Proteome/immunology, Proteome/isolation & purification, Proteomics/methods
Pubmed
Web of science
Open Access
Yes
Create date
26/07/2019 16:25
Last modification date
21/08/2019 5:35
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