Identification and validation of T-cell epitopes using the IFN-gamma ELISPOT assay

Methods Mol Biol. 2009;524:439-46. doi: 10.1007/978-1-59745-450-6_32.


The quest for new and enhanced vaccines requires improved means for identification of relevant antigens and the epitopes present within these. While CD8+ T cells recognize antigenic peptides presented in the context of MHC Class I, and play a key role in cellular immunity, CD4+ helper T-cells recognize antigens in the context of MHC Class II and assist other immune cells in orchestration of the defined immune response. Being a functional assay, ELISPOT can be used to define both MHC Class I and II epitopes as well as to validate them. Providing the quality of the cells used in the assay is sufficiently good, ELISPOT provides a solid platform for both screening and validating T-cell epitopes. While some ELISPOT assays rely upon loading of dendritic cells with antigen followed by incubation of HLA matched or autologous PBMC or enriched T cells, the assay presented here uses PBMC from diseased or vaccinated individuals to simultaneously identify and validate T-cell epitopes.

MeSH terms

  • CD4-Positive T-Lymphocytes / immunology
  • CD8-Positive T-Lymphocytes / immunology
  • Cells, Cultured
  • Epitope Mapping / methods*
  • Epitopes, T-Lymphocyte / analysis*
  • Epitopes, T-Lymphocyte / immunology
  • Humans
  • Immunoenzyme Techniques / methods*
  • Interferon-gamma / immunology*
  • Leukocytes, Mononuclear / immunology
  • Major Histocompatibility Complex


  • Epitopes, T-Lymphocyte
  • Interferon-gamma