Detection of Antigen Presentation by Murine Bone Marrow-Derived Dendritic Cells After Treatment with Nanoparticles

Methods Mol Biol. 2024:2789:161-169. doi: 10.1007/978-1-0716-3786-9_17.

Abstract

Nanoparticles are frequently considered in vaccine applications due to their ability to co-deliver multiple antigens and adjuvants to antigen-presenting cells. Some nanoparticles also have intrinsic adjuvant properties that further enhance their ability to stimulate immune cells. The delivery of tumor-specific antigens to antigen-presenting cells (APCs) with subsequent antigenic peptide presentation in the context of class I major histocompatibility complex (MHC-I) molecules represents an essential effort in developing nanotechnology-based cancer vaccines. Experimental models are, therefore, needed to gauge the efficiency of nanotechnology carriers in achieving peptide antigen delivery to APCs and presentation in the context of MHC-I. The assay described herein utilizes a model antigen ovalbumin and model APCs, murine bone marrow-derived dendritic cells. The 25-D1.16 antibody, specific to the ovalbumin (OVA) MHC-I peptide SIINFEKL, recognizes this peptide presented in the context of the murine H2-Kb class I MHC molecule, allowing the presentation of this antigen on APCs to be detected by flow cytometry after nanoparticle delivery.

Keywords: Antigen presentation; Dendritic cells; Nanoparticles; Vaccines.

MeSH terms

  • Animals
  • Antigen Presentation*
  • Antigens, Neoplasm
  • Bone Marrow
  • Dendritic Cells
  • Histocompatibility Antigens Class I
  • Mice
  • Mice, Inbred C57BL
  • Nanoparticles* / chemistry
  • Ovalbumin
  • Peptides

Substances

  • Ovalbumin
  • Histocompatibility Antigens Class I
  • Peptides
  • Antigens, Neoplasm