Development of a Versatile Cancer Vaccine Format Targeting Antigen-Presenting Cells Using Proximity-Based Sortase A-Mediated Ligation of T-Cell Epitopes

Bioconjug Chem. 2024 Nov 20;35(11):1805-1814. doi: 10.1021/acs.bioconjchem.4c00403. Epub 2024 Nov 7.

Abstract

Cancer vaccines are a promising strategy to increase tumor-specific immune responses in patients who do not adequately respond to checkpoint inhibitors. Cancer vaccines that contain patient-specific tumor antigens are most effective but also necessitate the production of patient-specific vaccines. This study aims to develop a versatile cancer vaccine format in which patient-specific tumor antigens can be site-specifically conjugated by a proximity-based Sortase A (SrtA)-mediated ligation (PBSL) approach to antibodies that specifically bind to antigen-presenting cells to stimulate immune responses. DEC205 and CD169 are both receptors expressed on antigen-presenting cells that can be targeted to deliver antigens and stimulate T-cell responses. We used the CRISPR/HDR platform to produce mouse heavy chain IgG2a antibodies with DEC205 or CD169 specificity containing an SrtA recognition motif followed by a SpyTag at the C-terminus. Using a recombinant protein of SrtA linked to SpyCatcher, we applied proximity-based SrtA-mediated ligation to ligate fluorescein isothiocyanate (FITC)-labeled or antigenic peptides to the antibodies. Ligated antibodies bound to DEC205-expressing dendritic cells or CD169-expressing macrophages both in vitro and in vivo. More importantly, immunization with DEC205- or CD169-specific Abs linked to T-cell epitopes efficiently stimulated T-cell responses in vivo. To conclude, we have developed a cancer vaccine format using PBSL that enables the rapid incorporation of tumor antigens and could potentially be implemented for the synthesis of personalized cancer vaccines.

MeSH terms

  • Aminoacyltransferases*
  • Animals
  • Antigen-Presenting Cells* / immunology
  • Antigens, Neoplasm / immunology
  • Bacterial Proteins* / immunology
  • Cancer Vaccines* / immunology
  • Cysteine Endopeptidases* / immunology
  • Epitopes, T-Lymphocyte* / immunology
  • Humans
  • Mice
  • Mice, Inbred C57BL

Substances

  • sortase A
  • Aminoacyltransferases
  • Cancer Vaccines
  • Cysteine Endopeptidases
  • Bacterial Proteins
  • Epitopes, T-Lymphocyte
  • Antigens, Neoplasm