Sensitive detection of human papillomavirus type 16 E7-specific T cells by ELISPOT after multiple in vitro stimulations of CD8+ T cells with peptide-pulsed autologous dendritic cells

Mol Cancer. 2006 Oct 26:5:49. doi: 10.1186/1476-4598-5-49.

Abstract

Background: Cervical cancer is the second most common gynecological cancer amongst women world-wide. Despite optimized protocols, standard treatments still face several disadvantages. Therefore, research aims at the development of immune-based strategies using tumor antigen-loaded dendritic cells for the induction of cellular anti-tumor immunity.

Results: In this study, we used dendritic cells loaded with the HLA-A2-restricted HPV type 16 E711-20 peptide in order to induce an in vitro CD8+ T cell response. For this purpose, peptide-pulsed dendritic cells were co-cultured with autologous CD8+ T cells. After 5 weekly stimulations with peptide-pulsed mature dendritic cells, cultured T cells were analyzed for antigen specificity by an IFN-gamma ELISPOT assay. Using this ELISPOT assay, we were able to detect E7-specific IFN-gamma-secreting CD8+ T cells in 5/5 healthy donors.

Conclusion: We show that peptide-pulsed mature dendritic cells are able to stimulate a HPV type 16 E7 peptide-specific immune response in vitro. These experiments describe an efficient culture protocol for antigen-specific T cells for use in pre-clinical vaccination research and confirm the need for sensitive T cell assays for detection of tumor-specific immune responses in vitro.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, Viral, Tumor / immunology*
  • CD8-Positive T-Lymphocytes / immunology*
  • Coculture Techniques
  • Dendritic Cells / immunology*
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Female
  • Humans
  • Immunophenotyping
  • Interferon-gamma / biosynthesis
  • K562 Cells
  • Oligopeptides / immunology
  • Oncogene Proteins, Viral / immunology*
  • Papillomavirus E7 Proteins
  • Papillomavirus Infections / immunology
  • Sensitivity and Specificity
  • Uterine Cervical Neoplasms / virology

Substances

  • Antigens, Viral, Tumor
  • Oligopeptides
  • Oncogene Proteins, Viral
  • Papillomavirus E7 Proteins
  • oncogene protein E7, Human papillomavirus type 16
  • Interferon-gamma