Identification and validation of T-cell epitopes using the IFN-gamma ELISPOT assay.
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.
Wulf
Markus
M
Thymed GmbH, Mikroforum Ring 2, 55234 Wendelsheim, Germany.
Hoehn
Petra
P
Trinder
Peter
P
D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002478:Cells, Cultured; D018604:Epitope Mapping / Q000379:methods*; D018984:Epitopes, T-Lymphocyte / Q000032:analysis* / Q000276:immunology; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods*; D007371:Interferon-gamma / Q000276:immunology*; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008285:Major Histocompatibility Complex
D018984:Epitopes, T-Lymphocyte; D007371:Interferon-gamma