Supplemental Figure 6 from T-cell Priming by High-Avidity Neoantigens in Lymph Nodes Augments Adoptive Immunotherapy

Supplemental Figure 6: Characterizing host immune cells in neoantigen versus wild-type tumors. (A) Breakdown of immune cells within B16KVP or B16F10 subcutaneous tumors (average taken of n=5). All mice were given either B16KVP or B16F10 tumors. Mice received TBI but no ACT. Five days later, mice were sacrificed and tumors processed for flow cytometry to evaluate immune cells present. Gated on Lymphocytes/Single Cells/Live/CD45+. Neutrophils = CD11b+ Ly6G+, Dendritic cells = CD11c+, macrophages/monocytes = CD11b+ Ly6C+, CD4 T cells = CD3+ CD4+, CD8 T cells = CD3+ CD8+, B cells = CD19+, and NK cells = NK1.1+. (B) Breakdown of immune cells within B16KVP or B16F10 subcutaneous tumors that received naïve pmel-1 ACT in addition to TBI. Same gating used as A. (C) Percentage of immune cells (CD45+) in B16KVP or B16F10 tumors six days after 5Gy TBI. Kruskal-Wallis test with multiple comparisons performed. (* denotes p < 0.05). (D) Percentage of host CD8+ T cells present in each tumor type +/- ACT. Gated on Lymphocytes/Single Cells/Live/CD45+. Kruskal-Wallis test with multiple comparisons performed. NS denotes p > 0.05. (E) Percentage of trp-2 tetramer positive host T cells present in the spleen, blood, and LN in long-term survivor mice 100+ days post-ACT. Gated on Lymphocytes/Live/Single Cells/CD45+/Thy1.1-Thy1.2+.