title: Mapping the HLA ligandome landscape of acute myeloid leukemia: a targeted approach toward peptide-based immunotherapy.
abstract: Identification of physiologically relevant peptide vaccine targets calls for the direct analysis of the entirety of naturally presented human leukocyte antigen (HLA) ligands, termed the HLA ligandome. In this study, we implemented this direct approach using immunoprecipitation and mass spectrometry to define acute myeloid leukemia (AML)-associated peptide vaccine targets. Mapping the HLA class I ligandomes of 15 AML patients and 35 healthy controls, more than 25 000 different naturally presented HLA ligands were identified. Target prioritization based on AML exclusivity and high presentation frequency in the AML cohort identified a panel of 132 LiTAAs (ligandome-derived tumor-associated antigens), and 341 corresponding HLA ligands (LiTAPs (ligandome-derived tumor-associated peptides)) represented subset independently in >20% of AML patients. Functional characterization of LiTAPs by interferon-γ ELISPOT (Enzyme-Linked ImmunoSpot) and intracellular cytokine staining confirmed AML-specific CD8(+) T-cell recognition. Of note, our platform identified HLA ligands representing several established AML-associated antigens (e.g. NPM1, MAGED1, PRTN3, MPO, WT1), but found 80% of them to be also represented in healthy control samples. Mapping of HLA class II ligandomes provided additional CD4(+) T-cell epitopes and potentially synergistic embedded HLA ligands, allowing for complementation of a multipeptide vaccine for the immunotherapy of AML.
authors.affiliation: 1] Department of Immunology, Institute for Cell Biology, University of Tübingen, Tübingen, Germany [2] Department of Hematology and Oncology, University of Tübingen, Tübingen, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Berlin
authors.affiliation: Department of Immunology, Institute for Cell Biology, University of Tübingen, Tübingen, Germany.
authors.forename: D J
authors.initials: DJ
authors.lastname: Kowalewski
authors.affiliation: Department of Immunology, Institute for Cell Biology, University of Tübingen, Tübingen, Germany.
authors.forename: H
authors.initials: H
authors.lastname: Schuster
authors.affiliation: 1] Department of Immunology, Institute for Cell Biology, University of Tübingen, Tübingen, Germany [2] Department of Hematology and Oncology, University of Tübingen, Tübingen, Germany.
authors.forename: N
authors.initials: N
authors.lastname: Mirza
authors.affiliation: 1] Department of Immunology, Institute for Cell Biology, University of Tübingen, Tübingen, Germany [2] Department of Hematology and Oncology, University of Tübingen, Tübingen, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Walz
authors.affiliation: Hospital Group South-West, Department of Orthopedics, Calw, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Handel
authors.affiliation: Institute for Clinical and Experimental Transfusion Medicine, University of Tübingen, Tübingen, Germany.
authors.forename: B
authors.initials: B
authors.lastname: Schmid-Horch
authors.affiliation: 1] Department of Hematology and Oncology, University of Tübingen, Tübingen, Germany [2] Clinical Collaboration Unit Translational Immunology, German Cancer Consortium (DKTK) and German Cancer Research Center (DKFZ), Heidelberg, Germany.
authors.forename: H R
authors.initials: HR
authors.lastname: Salih
authors.affiliation: Department of Hematology and Oncology, University of Tübingen, Tübingen, Germany.
authors.forename: L
authors.initials: L
authors.lastname: Kanz
authors.affiliation: 1] Department of Immunology, Institute for Cell Biology, University of Tübingen, Tübingen, Germany [2] Clinical Collaboration Unit Translational Immunology, German Cancer Consortium (DKTK) and German Cancer Research Center (DKFZ), Heidelberg, Germany.
authors.forename: H-G
authors.initials: HG
authors.lastname: Rammensee
authors.affiliation: 1] Department of Immunology, Institute for Cell Biology, University of Tübingen, Tübingen, Germany [2] Clinical Collaboration Unit Translational Immunology, German Cancer Consortium (DKTK) and German Cancer Research Center (DKFZ), Heidelberg, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Stevanović
authors.affiliation: 1] Department of Immunology, Institute for Cell Biology, University of Tübingen, Tübingen, Germany [2] Department of Hematology and Oncology, University of Tübingen, Tübingen, Germany.
authors.forename: J S
authors.initials: JS
authors.lastname: Stickel
mesh_terms: D000595:Amino Acid Sequence; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D019496:Cancer Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D016022:Case-Control Studies; D018984:Epitopes, T-Lymphocyte / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D015870:Gene Expression; D015395:Histocompatibility Antigens Class I / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D000949:Histocompatibility Antigens Class II / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D006801:Humans; D047468:Immunoprecipitation; D016233:Immunotherapy, Active / Q000379:methods*; D015470:Leukemia, Myeloid, Acute / Q000235:genetics / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D008024:Ligands; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000235:genetics / Q000276:immunology*; D000090243:Nucleophosmin; D010449:Peptide Mapping; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
chemical_list: D019496:Cancer Vaccines; D018984:Epitopes, T-Lymphocyte; D015395:Histocompatibility Antigens Class I; D000949:Histocompatibility Antigens Class II; D008024:Ligands; C000716967:NPM1 protein, human; D009363:Neoplasm Proteins; D010455:Peptides; D000090243:Nucleophosmin
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