Simultaneous Identification of Functional Antigen-Specific CD8⁺ and CD4⁺ Cells after In Vitro Expansion Using Elongated Peptides.

ABSTRACT: Elongated peptides (EPs), containing possibly one or multiple epitope/s, are increasingly used for the screening of antigen-specific CD8⁺ and CD4⁺ cell responses. Here, we present an in vitro protocol that allows the amplification of antigen-specific cells and the subsequent functional analysis of both T cell types using EPs. Known viral-derived epitopes were elongated to 20 mer EPs on the N-, C-, and both termini for HLA class I binders, or on the N- and C- termini for HLA class II binders. With EP stimulation only, the percentage of responding CD8⁺ T cells was dependent on the elongation site of the EP, whereas CD4⁺ T cell responses were completely lost in 22% of the tests performed ex vivo. A short-term amplification step plus the addition of

SUBMITTER: Schuhmacher J 

PROVIDER: S-EPMC9654851 | biostudies-literature | 2022 Oct

REPOSITORIES: biostudies-literature