Critical components of a DNA fusion vaccine able to induce protective cytotoxic T cells against a single epitope of a tumor antigen

J Immunol. 2002 Oct 1;169(7):3908-13. doi: 10.4049/jimmunol.169.7.3908.

Abstract

DNA vaccines can activate immunity against tumor Ags expressed as MHC class I-associated peptides. However, priming of CD8(+) CTL against weak tumor Ags may require adjuvant molecules. We have used a pathogen-derived sequence from tetanus toxin (fragment C (FrC)) fused to tumor Ag sequences to promote Ab and CD4(+) T cell responses. For induction of CD8(+) T cell responses, the FrC sequence has been engineered to remove potentially competitive MHC class I-binding epitopes and to improve presentation of tumor epitopes. The colon carcinoma CT26 expresses an endogenous retroviral gene product, gp70, containing a known H2-L(d)-restricted epitope (AH1). A DNA vaccine encoding gp70 alone was a poor inducer of CTL, and performance was not significantly improved by fusion of full-length FrC. However, use of a minimized domain of FrC, with the AH1 sequence fused to the 3' position, led to rapid induction of high levels of CTL. IFN-gamma-producing epitope-specific CTL were detectable ex vivo and these killed CT26 targets in vitro. The single epitope vaccine was more effective than GM-CSF-transfected CT26 tumor cells in inducing an AH1-specific CTL response and equally effective in providing protection against tumor challenge. Levels of AH1-specific CTL in vivo were increased following injection of tumor cells, and CTL expanded in vitro were able to kill CT26 cells in tumor bearers. Pre-existing immunity to tetanus toxoid had no effect on the induction of AH1-specific CTL. These data demonstrate the power of epitope-specific CTL against tumor cells and illustrate a strategy for priming immunity via a dual component DNA vaccine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Neoplasm / administration & dosage
  • Antigens, Neoplasm / genetics
  • Antigens, Neoplasm / immunology*
  • Cancer Vaccines / administration & dosage
  • Cancer Vaccines / chemical synthesis
  • Cancer Vaccines / genetics
  • Cancer Vaccines / immunology*
  • Colonic Neoplasms / immunology
  • Colonic Neoplasms / pathology
  • Colonic Neoplasms / prevention & control
  • Cytotoxicity, Immunologic* / genetics
  • Epitopes, T-Lymphocyte / administration & dosage
  • Epitopes, T-Lymphocyte / genetics
  • Epitopes, T-Lymphocyte / immunology*
  • Growth Inhibitors / administration & dosage
  • Growth Inhibitors / chemical synthesis
  • Growth Inhibitors / genetics
  • Growth Inhibitors / immunology
  • Injections, Intramuscular
  • Interferon-gamma / biosynthesis
  • Lymphocyte Activation* / genetics
  • Mice
  • Mice, Inbred BALB C
  • Neoplasm Transplantation
  • Recombinant Fusion Proteins / administration & dosage
  • Recombinant Fusion Proteins / chemical synthesis
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology*
  • Retroviridae Proteins, Oncogenic / administration & dosage
  • Retroviridae Proteins, Oncogenic / genetics
  • Retroviridae Proteins, Oncogenic / immunology
  • T-Lymphocytes, Cytotoxic / immunology*
  • T-Lymphocytes, Cytotoxic / metabolism
  • T-Lymphocytes, Cytotoxic / transplantation
  • Tetanus Toxoid / administration & dosage
  • Tetanus Toxoid / immunology
  • Vaccines, DNA / administration & dosage
  • Vaccines, DNA / chemical synthesis
  • Vaccines, DNA / genetics
  • Vaccines, DNA / immunology*
  • Viral Envelope Proteins / administration & dosage
  • Viral Envelope Proteins / genetics
  • Viral Envelope Proteins / immunology

Substances

  • Antigens, Neoplasm
  • Cancer Vaccines
  • Epitopes, T-Lymphocyte
  • Growth Inhibitors
  • Recombinant Fusion Proteins
  • Retroviridae Proteins, Oncogenic
  • Tetanus Toxoid
  • Vaccines, DNA
  • Viral Envelope Proteins
  • Interferon-gamma