Fig. 3.

Pep R in combination with anti-PD-1 increased Granzyme B infiltration and reduced Treg recruitment in B16-hCXCR4 tumors. Granzyme B infiltration and FoxP3 positive immune cells was assessed by immunohistochemistry in tumors from B16-hCXCR4 (a-d). a Representative microphotographs of Granzyme B across treatment groups. Granzyme B staining was mainly detected in cytoplasmic granules of cytolytic T-lymphocytes and natural killer cells (red staining). Counterstaining of nuclei by hematoxilyn (blue) (magnification 400x). b. GZMB was quantified with the AxioVision Imaging System version 4.8 microscope expressed as IHC positive cell/mm². Untreated mice: 3.23 ± 1.01 (n = 8); anti-PD-1 = 7.02 ± 1.19 (n = 8); Pep R = 7.44 ± 1.52 (n = 8); anti-PD-1 + Pep R = 15.6 ± 3.60 (n = 7). c Representative microphotographs of FoxP3. FoxP3 staining showed nuclear immunoreactivity in lymphocytes (red staining, magnification 400x). d Quantification FoxP3 was expressed as IHC positive cell/mm². Data are shown as mean ± SEM. Untreated mice: 8.70 ± 2.17 (n = 5); anti-PD-1 = 10.18 ± 0.79 (n = 6); Pep R = 7.65 ± 3.31 (n = 8); anti-PD-1 + Pep R = 4.08 ± 1.70 (n = 6). ANOVA Posthoc Tukey HSD. * P < 0.05