High-sensitivity HLA class I peptidome analysis enables a precise definition of peptide motifs and the identification of peptides from cell lines and patients’ sera
| Autor: | Benjamin Weide, Dario Neri, Danilo Ritz, Tim Fugmann, Andreas Gloger, Claus Garbe |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Amino Acid Motifs HL-60 Cells Peptide Human leukocyte antigen Computational biology Biochemistry Article 03 medical and health sciences 0302 clinical medicine Antigen Antigens Neoplasm Consensus Sequence Biomarkers Tumor Consensus sequence Humans Protein Interaction Domains and Motifs Spondylitis Ankylosing Multiplex Amino Acid Sequence Melanoma Molecular Biology Peptide sequence chemistry.chemical_classification biology Histocompatibility Antigens Class I 3. Good health HEK293 Cells 030104 developmental biology chemistry Case-Control Studies Immunology biology.protein Antibody Peptides CD8 Protein Binding 030215 immunology |
| Zdroj: | PROTEOMICS |
| ISSN: | 1615-9853 |
| DOI: | 10.1002/pmic.201500445 |
| Popis: | The characterization of peptides bound to human leukocyte antigen (HLA) class I is of fundamental importance for understanding CD8+ T cell-driven immunological processes and for the development of immunomodulatory therapeutic strategies. However, until now, the mass spectrometric analysis of HLA-bound peptides has typically required billions of cells, still resulting in relatively few high-confidence peptide identifications. Capitalizing on the recent developments in mass spectrometry and bioinformatics, we have implemented a methodology for the efficient recovery of acid-eluted HLA peptides after purification with the pan-reactive antibody W6/32 and have identified a total of 27 862 unique peptides with high confidence (1% false discovery rate) from five human cancer cell lines. More than 93% of the identified peptides were eight to 11 amino acids in length and contained signatures that were in excellent agreement with published HLA binding motifs. Furthermore, by purifying soluble HLA class I complexes (sHLA) from sera of melanoma patients, up to 972 high-confidence peptides could be identified, including melanoma-associated antigens already described in the literature. Knowledge of the HLA class I peptidome should facilitate multiplex tetramer technology-based characterization of T cells, and allow the development of patient selection, stratification and immunomodulatory therapeutic strategies. |
| Databáze: | OpenAIRE |
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