Figure 2.

GVAX Vaccination Induces a Systemic Cytotoxic CD8 T Cell Response to Rat-Neu Neoantigen.

(a) Treatment scheme for the neu-expressing (GM-CSF–secreting) vaccination (GVAX) group. One day after the vaccination, 1 × 10⁶ high-avidity CFSE-labeled Thy1.2⁺ RNEU_420–429-specific CD8 T cells were adoptively transferred (AT) into mice. On day 5, inguinal LNs (ILNs) and spleens were harvested and analyzed by flow cytometry. (b) Gating strategy to profile RNEU-specific CD8 T cells by flow cytometry. FVB/N-derived T cell clone TCRVβ4 was gated based on CD45⁺CD8⁺Thy1.2⁺. (c) Percentages of proliferating TCRVβ4⁺ CD8 T cells in indicated tissues (representative flow plots and quantification; n ≥ 4 per group, repeated x 2). (d) Percentages of cytokine production by RNEU-specific TCRVβ4⁺ CD8 T cells (representative flow plots and quantification; n ≥ 4 per group, repeated x 2). (e) Percentages of polyfunctional RNEU-specific TCRVβ4⁺ IFNγ⁺GzB⁺TNFα⁺ CD8 T cells in indicated tissues (n ≥ 4 per group, repeated x 2). Proliferating TCRVβ4⁺ CD8 T cells, and their cytokine production, were calculated as fraction of TCRVβ4⁺ CD8 T cells.