Fig 1: Conjugation of peptide antigens to pAntp enhances vaccine potency without skewing T cell phenotypes, leading to robust expansion of therapeutically effective T cells. (A) Timeline of immunization experiments. (B) Percentage of IFN-?+ of CD8+ T cells (or CD4+ cells for M30 and M48) after a prime and boost using the indicated long antigen peptide or antigen-pAntp, along with c-di-GMP adjuvant, as compared to the respective antigen. (C and D) C57BL C57BL/6 mice (n = 5 animals per group) were immunized with 5 nmol Adpgk or Adpgk-pAntp peptide combined with 25 µg cyclic-di-GMP on days 0 and 14, and then on day 21 PBMCs were restimulated with optimal Adpgk peptide ex vivo and stained for flow cytometry analysis. Shown are representative flow cytometry plots, percentages, and quantification of: (C) CD8+/IFN-?+/IL-7Rhi/KLRG1lo MPECs, CD8+/IFN-?+/IL-7Rlo/KLRG1hi SLECs, and CD8+/IFN-?+/IL-7Rlo/KLRG1lo early effectors (EE); (D) PD-1+/TCF-1+ expression. Statistical analyses for C and D were performed using a two-way ANOVA with Šídák’s multiple comparisons test. (E) Timeline for therapeutic efficacy study using the MC-38 tumor model. (F) Corresponding tumor growth curves with a two-way ANOVA statistical comparison followed by Tukey posttest of untreated control to Adgpk-pAntp. (G) overall survival curves, analyzed with the Mantel–Cox log-rank test. For all sections ****P < 0.0001; ***P < 0.001; **P < 0.01, *P < 0.05; n.s., not significant.