Identification of human MHC class I binding peptides using the iTOPIA- epitope discovery system.

CD8+ T cells recognize antigenic peptides presented in the context of MHC class I. They play a key role in cellular immunity and are crucial for longterm protective immunity to many infectious diseases. The quest for new and enhanced vaccines requires improved means for identification of relevant antigens and the epitopes present within these. While there are several algorithms available for epitope prediction (all of which work to differing degrees of success), the definition of actual MHC class I-binding epitopes is very reliant on time-consuming and difficult to perform functional assays using often very limited biological material. The iTOPIA assay is quick and easy to perform and determines real-binding to MHC class I molecules. It provides an excellent platform for screening and elimination of potential epitopes and identification of novel epitopes prior to validation with a relevant functional assay.

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.