Figure 6.

The combination of G with P and H promotes M1 polarization and T cell activation in the KPC-Luc-TME

(A) t-SNE plot analysis showed ten subset clusters representing five major cell types (M1 and M2 macrophages, monocytes, granulocytes, and dendritic cells).

(B) Well-established gene markers recognized subclusters for macrophages.

(C and D) GPH treatment increases the percentage of M1 macrophages polarization, monocytes, and granulocytes.

(E) The percentages of CD45⁺ CD11b⁺ Ly6C⁻ Ly6G⁻ F4/80+ macrophage, CD11b⁺ Ly6G⁺ Ly6C⁺ gMDSC, CD11b⁺ Ly6G⁻ Ly6C^hi mMDSC, CD11b⁻ CD11c^hi MHCII^hi XCR1⁺ cDC1, CD11b⁺ CD11c^hi MHCII^hi CD172α⁺ cDC2, and CD11b⁻ CD11c^int pDC subsets were evaluated in sham, PH, G, and GPH (N = 4 mice/group). Data are representative of a single experiment. One-way ANOVA run, followed by Tukey’s multiple comparisons test, determined p values. Error bars indicate SD.

(F) Dot plot showed that GPH treatment modulate the myeloid population representing gene markers. GPH treatment decreases the expression of M2 markers and increases dendritic cell (DC) markers.

(G) The percentages of macrophages CD45⁺ CD11b⁺ Ly6G⁻ Ly6C⁻ F4/80⁺ macrophage and median fluorescent intensity of MHCII and CD206 were evaluated in sham, PH, G, and GPH (N = 4 mice/group). Data are representative of a single experiment. One-way ANOVA run, followed by Tukey’s multiple comparisons test, determined p values. Error bars indicate SD.

(H and I) Well-established gene markers recognized subclusters for T cells.

(J) t-SNE plot showed that GPH treatment decreases the percentage of Cd4 and Cd8 Tregs and increases the percentage of Cd4⁺ and Cd8⁺ T cells as well as NK cells.

(K) Proportions of CD4⁺ Foxp3⁻, CD4⁺ Foxp3⁺, CD8⁺ T cells, γδ T cells, and NK cells in KPC-Luc-TME were evaluated in sham, PH, G, and GPH (N = 4 mice/group). Data are representative of a single experiment. One-way ANOVA run, followed by Tukey’s multiple comparisons test, determined p values.

(L–O) Dot plot showed that GPH treatment increases the expression of the Cd4, Cd3e, Nkg7, Ctla-4, and Pd-1 as well as Ifnγ, Prf1, and Tbx21 and decreases Foxp3. CD4⁺ FOXP3⁺ (M), CD4⁺ Foxp3⁻ (N), and CD8⁺ T cell (O) subsets from KPC-Luc-TME were graphed as mean fluorescent intensity per mg tumor weight. Data are representative of a single experiment. Welch’s t test determined p values; Asterisks (∗p < 0.01, ∗∗p < 0.001, ∗∗∗p < 0.0001, and ∗∗∗∗p < 0.00001) denote the level of significance (between the sham and treatment groups), ns denote non-significant. Error bars indicate SD. See also Figures S9–S11.