Engineering Cell Lines for Specific Human Leukocyte Antigen Presentation

Methods Mol Biol. 2023:2691:351-369. doi: 10.1007/978-1-0716-3331-1_25.

Abstract

Epitope-specific immunotherapies have enabled the targeted treatment of a variety of diseases, ranging from cancer, infection, and autoimmune disorders. For CD8+ T cell-based therapies, the precise identification of immunogenic peptides presented by human leukocyte antigen (HLA) class I is essential which can be achieved by immunopeptidomics. Here, using lentivirus-mediated transduction and cell sorting approaches, we present a method to engineer a cell line that does not express its native HLA but instead expresses an HLA of interest (in this instance HLA-A*02:01). This technique can be used to elucidate the immunopeptidome of cell lines expressing different HLAs.

Keywords: Cancer; Cell lines; Gene editing; HLA; Immunopeptidomics; Immunotherapy; Lentivirus; Neoantigens; Transduction.

MeSH terms

  • Antigen Presentation
  • Antigens, Neoplasm*
  • Cell Line, Tumor
  • Epitopes, T-Lymphocyte
  • HLA Antigens / genetics
  • Histocompatibility Antigens Class I* / genetics
  • Histocompatibility Antigens Class II
  • Humans

Substances

  • Antigens, Neoplasm
  • Histocompatibility Antigens Class I
  • HLA Antigens
  • Histocompatibility Antigens Class II
  • Epitopes, T-Lymphocyte