Fig. 3.

Ex vivo responses against several TGFβ1-derived epitopes. A Amplitude of responses in PBMCs from both cancer patients and healthy donor PBMCs in ex vivo ELISPOT. PBMCs were rested overnight after thawing and then plated directly in ELISPOT wells and stimulated with epitopes for 48 H in ELISPOT wells. The results shown demonstrate a response against TGFβ-05, TGFβ-15, and TGFβ-26. B Examples of ex vivo ELISPOT responses against several of the TGFβ1 lead epitopes. C CD8⁺ T-cell response identified against the epitope TGFβ-15 after only five H of stimulation using ICS in an ex vivo setting. D PBMCs from a patient with prostate cancer displaying a CD8⁺ T-cell response against the TGFβ-15 epitope after 18 H stimulation with the epitope with a prior 14 days of in vitro stimulation. E Gate set on CD4⁺ T cells from a TGFβ-15-specific T-cell culture from donor UR1121.14 cells stimulated with TGFβ-15. Cells were enriched twice using the MACS CD137 enrichment method as described in the Materials and Methods. F Gate set on CD8⁺ T cells in the TGFβ-15-specific T-cell culture described above