High-throughput and Sensitive Immunopeptidomics Platform Reveals Profound Interferonγ-Mediated Remodeling of the Human Leukocyte Antigen (HLA) Ligandome.
- Chong C From the ‡Ludwig Institute for Cancer Research, University of Lausanne, 1066 Epalinges, Switzerland.
- Marino F From the ‡Ludwig Institute for Cancer Research, University of Lausanne, 1066 Epalinges, Switzerland.
- Pak H From the ‡Ludwig Institute for Cancer Research, University of Lausanne, 1066 Epalinges, Switzerland.
- Racle J From the ‡Ludwig Institute for Cancer Research, University of Lausanne, 1066 Epalinges, Switzerland.
- Daniel RT ¶Service of Neurosurgery, University Hospital of Lausanne, 1011 Lausanne, Switzerland.
- Müller M ‖Vital IT, Swiss Institute of Bioinformatics, 1015 Lausanne, Switzerland.
- Gfeller D From the ‡Ludwig Institute for Cancer Research, University of Lausanne, 1066 Epalinges, Switzerland.
- Coukos G From the ‡Ludwig Institute for Cancer Research, University of Lausanne, 1066 Epalinges, Switzerland.
- Bassani-Sternberg M From the ‡Ludwig Institute for Cancer Research, University of Lausanne, 1066 Epalinges, Switzerland; Michal.bassani@chuv.ch.
- 2017-12-16
Published in:
- Molecular & cellular proteomics : MCP. - 2018
B-Lymphocytes
Cell Line
Histocompatibility Antigens Class I
Histocompatibility Antigens Class II
Humans
Interferon-gamma
Ligands
Neoplasms
Peptides
Proteomics
T-Lymphocytes
English
Comprehensive knowledge of the human leukocyte antigen (HLA) class-I and class-II peptides presented to T-cells is crucial for designing innovative therapeutics against cancer and other diseases. However methodologies for their purification for mass-spectrometry analysis have been a major limitation. We designed a novel high-throughput, reproducible and sensitive method for sequential immuno-affinity purification of HLA-I and -II peptides from up to 96 samples in a plate format, suitable for both cell lines and tissues. Our methodology drastically reduces sample-handling and can be completed within five hours. We challenged our methodology by extracting HLA peptides from multiple replicates of tissues (n = 7) and cell lines (n = 21, 108 cells per replicate), which resulted in unprecedented depth, sensitivity and high reproducibility (Pearson correlations up to 0.98 and 0.97 for HLA-I and HLA-II). Because of the method's achieved sensitivity, even single measurements of peptides purified from 107 B-cells resulted in the identification of more than 1700 HLA-I and 2200 HLA-II peptides. We demonstrate the feasibility of performing drug-screening by using ovarian cancer cells treated with interferon gamma (IFNγ). Our analysis revealed an augmented presentation of chymotryptic-like and longer ligands associated with IFNγ induced changes of the antigen processing and presentation machinery. This straightforward method is applicable for basic and clinical applications.
- Language
-
- English
- Open access status
- hybrid
- Identifiers
-
- DOI 10.1074/mcp.TIR117.000383
- PMID 29242379
- Persistent URL
- https://folia.unifr.ch/global/documents/259373
Statistics
Document views: 46
File downloads:
- Full-text: 0