Fig. 3.

HA29- and HI12-scDbs induce polyfunctional CD8⁺ T cell responses. T2 cells were pulsed with the ALT or ILK peptides at the indicated concentrations. Next, 10 × 10⁴ peptide-pulsed cells were cocultured with 5 × 10⁴ preactivated CD8⁺T cells (1:2 E:T) in the presence of 0.25 nM HA29- or HI12-scDb for 72 h. Supernatants were assayed with Legendplex or MIP1β ELISAs. Specific activation of CD8⁺T cells by (A) HA29 or (B) HI12 was observed by the secretion of Granzyme A (A and B, Top) and IFN-γ (A and B, Bottom) in response to cells pulsed with the cognate but not irrelevant peptide. Data indicate mean ± range of two biological replicates. Similar specific responses were also observed using assays for other effector molecules (SI Appendix, Fig. S8).