Proteasomal targeting and minigene repetition improve cell-surface presentation of a transfected, modified melanoma tumour antigen

Scand J Immunol. 2004 Feb;59(2):220-7. doi: 10.1111/j.0300-9475.2004.01374.x.

Abstract

Melanoma antigen recognized by T cell 1 (MART-1) is regarded as a candidate peptide for vaccination against malignant melanoma, and it is of importance to develop strategies to improve the vaccine-elicited T-cell activation towards MART-1. T-cell activation is, among other determinants, dependent on the density of specific major histocompatibility complex-peptide complexes on the surface of the antigen-presenting cell. In this study, we explored the cell-surface presentation of a substituted MART-1 peptide encoded by transfected minigenes. We investigated the potential of proteasomal targeting compared to non-proteasomal targeting of the epitope to increase its cell-surface presentation. Furthermore, we explored the potential of incorporating multiple minigenes instead of one to increase cell-surface presentation. We show that both proteasomal targeting and repetition of the minigene increase cell-surface presentation of the epitope and propose both these approaches as potential strategies in DNA vaccines to increase MART-1-specific T-cell activation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Antigen Presentation / genetics
  • Antigen Presentation / immunology
  • Antigens, Neoplasm
  • Blotting, Western
  • Cancer Vaccines / genetics
  • Cancer Vaccines / immunology*
  • Cell Line
  • Cysteine Endopeptidases / genetics
  • Cysteine Endopeptidases / immunology*
  • Cytotoxicity Tests, Immunologic
  • Epitopes / immunology
  • Humans
  • MART-1 Antigen
  • Melanoma / immunology*
  • Melanoma / therapy
  • Molecular Sequence Data
  • Multienzyme Complexes / genetics
  • Multienzyme Complexes / immunology*
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / immunology*
  • Plasmids / genetics
  • Plasmids / immunology
  • Polymerase Chain Reaction
  • Proteasome Endopeptidase Complex
  • Repetitive Sequences, Nucleic Acid
  • T-Lymphocytes / immunology
  • Transfection
  • Vaccines, DNA / immunology

Substances

  • Antigens, Neoplasm
  • Cancer Vaccines
  • Epitopes
  • MART-1 Antigen
  • MLANA protein, human
  • Multienzyme Complexes
  • Neoplasm Proteins
  • Vaccines, DNA
  • Cysteine Endopeptidases
  • Proteasome Endopeptidase Complex