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. 2025 Aug 5;99(9):e00188-25. doi: 10.1128/jvi.00188-25

Fig 4.

Box plot and heat maps depict methylation status across cytokine, cytolytic, and naïve markers. IST group shows methylation loci at LEF1, TCF7, and demethylation at IFNG, GZMB, and PRF1 sites, consistent with enhanced transcriptional potential.

Epigenetic signatures of MART-1-IST treatment are consistent with differentiated cells. (A) T cell multipotency index. T cell multipotency score was generated from 0 to 1 for each indicated sample (UN, untreated; DC, dendritic cell treated; and IST, MART-1 Immuno-STAT treated) and compared to true naive (Tnaive, 1) and terminally exhausted (HIV_EC, 0) reference samples. Each dot represents one technical replicate from two donors shown as triangles or squares. (B) Normalized plots of CpG methylation from donor HGLK005 at sites surrounding and within differentially methylated regions of effector molecules (IFNG, TNF, GZMB, and PRF1) and (C) naive associated transcription factors (LEF1 and TCF7) obtained from EM Seq analysis. Red and blue lines depict methylated and unmethylated CpG sites, respectively, while green lines depict differentially methylated regions between IST and mDC. Significance was calculated by ordinary one-way ANOVA followed by Tukey’s multiple comparisons test using GraphPad Prism 10.4.0.