Figure 2 from Epigenetic Induction of Cancer-Testis Antigens and Endogenous Retroviruses at Single-Cell Level Enhances Immune Recognition and Response in Glioma

DAC immunesensitization promotes specific, robust, and polyfunctional NY-ESO-1 TCR CD8⁺ T-cell responses in primary GBM cell lines. A, Representative Bis-Seq-TA cloning lollipop diagram and (B) percent of methylated CpGs to total CpGs of first eight sites of analyzed CpG island in primary GS277 cells treated with DMSO or 0.5 µmol/L DAC for 12 days. C, Average fold change of mRNA expression of treated GS277 (n = 3; **, P < 0.01; two-tailed unpaired t test). D, Cytotoxic analysis of treated GS277 over a 40-hour period when co-incubated with NY-ESO-1 TCR-transduced effector T cells at E:T ratio of 1:1 and (E) at multiple E:T ratios (1:5, 1:2, 1:1, 2:1) at 20-hour (n = 3; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; multiple unpaired t tests). F, Uniform manifold approximation projection (UMAP) of CD8⁺ T cells transduced with NY-ESO-1 TCR CD8 or untransduced control CD8 (CTR) from four stimulated groups: NY-ESO-1 CD8⁺ alone (n = 1665 CD8⁺ cells), NY-ESO-1 CD8⁺ stimulated with DBTRG-05 (n = 1902 CD8⁺ cells), NY-ESO-1 CD8⁺ stimulated with DBTRG-05 + DAC (n = 284 CD8⁺ cells), CTR CD8 stimulated with DBTRG-05 + DAC (n = 807 CD8⁺ cells). G and H, PSI defined as the percentage of polyfunctional cells, multiplied by MFI of the proteins secreted by the polyfunctional cells for stimulated CD8⁺ at the single-cell resolution across DBTRG-05 co-cultures.