PMC

(A) Expression of PD-L1 on M05 tumor cells after (left) no treatment or (right) IL PV-10 injection as measured by flow cytometry. Gray histogram: isotype control. (B) Expression of PD-1 on total CD3⁺ and CD3⁺CD8⁺ tumor infiltrating lymphocytes (TIL) isolated from untreated or IL PV-10 treated M05 tumors.

Mice received 3x10⁵ M05 tumor cells SC on a single flank on Day 0. On Day 7, mice received either 50 μl PV-10 or PBS IL, and mice received anti-PD-1 antibody IP twice per week beginning on Day 8. Tumors were measured until the endpoint was reached.

Mice received 1x10⁵ B16 tumor cells SC on bilateral flanks on Day 0. On Day 7, right flank tumors were injected with 50 μl PV-10 or PBS IL. Mice received 300 μg anti-PD-L1 or NrIgG antibodies beginning on Day 8 and continuing 2x/week until mice had reached endpoint. Combination of PV-10 and anti-PD-L1 led to a delayed tumor growth in A) B16 tumors treated with IL PV-10 and B) untreated B16 bystander tumors.

Mice received 3x10⁵ M05 tumor cells SC on a both flanks on Day 0. On Day 7, mice received 50 μl PV-10 or PBS IL, and mice received anti-PD-L1 or IgG antibody IP twice per week beginning on Day 8. Mice were euthanized on Day 21, tumors were harvested and TILs were isolated from tumors. Infiltrating (A) CD4⁺ and (B) CD8⁺ T cells were measured by flow cytometry.

Mice received 3x10⁵ M05 tumor cells SC on a single flank on Day 0. On Day 7, mice received 50 μl PV-10 or PBS IL, and mice received anti-PD-1 antibody IP twice per week beginning on Day 8. Spleens were harvested on Day 21–24. (A) Splenocytes were co-cultured with M05 cells for 48 hours and supernatants were collected. IFN-gamma was measured by ELISA. (B) CD8⁺ and OVA-tetramer positive T cells were measured by flow cytometry.

Mice received 3x10⁵ M05 tumor cells SC on a single flank on Day 0, and were given 300 μg IP NrIgG control antibodies, 2.43 antibody to deplete CD8⁺ T cells, GK1.5 antibody to deplete CD4⁺ T cells, or PC61 antibody to deplete CD25⁺ Tregs. Antibodies were given twice per week until the completion of the experiment. On Day 7, mice received 50 μl PV-10 or PBS IL, and mice received anti-PD-1 antibody IP twice per week beginning on Day 8.

Mice received 3x10⁵ M05 tumor cells SC on bilateral flanks on Day 0. On Day 7, right flank tumors were injected with 50 μl PV-10 or PBS IL. Mice received 300 μg anti-PD-1 and NrIgG antibodies beginning on Day 8 and continuing 2x/week until mice had reached endpoint. Tumor growth was measured in (left panel) tumors treated with IL PBS or PV-10 and (right panel) untreated, bystander tumors. Each line represents the growth of tumor in a single mouse (n = 5–6 mice per treatment group).