Figure 1. Chronological order of experiments for the identification of Flu-specific clones for BC-143. (A) T-cell responses measured by IFN-γ ELISPOT assay: CD8⁺ T-cell responses to Flu-M- and SSX-2-peptides (left) as well as CD4⁺ T-cell response to Flu-NP- and SSX-2-peptides (right). Five × 10⁴ (upper part) and 1 × 10⁴ (lower part) cells per well were used. (B) T-cell responses measured by IFN-γ-secretion assay: CD4⁺ T-cell responses to Flu-NP and SSX-2 peptides by IFN-γ secretion (upper part) and CD8⁺ T-cell responses to Flu-M- and SSX-2-peptides measured by a combination of IFN-γ secretion and tetramer staining (lower part). (C) Clone specificity analysis: Analysis of CD8⁺ T-cell clone specificity via IFN-γ ELISPOT (upper part) and tetramer-FACS analysis (lower part).