Fig. 7: VSV-SB-HCC1,2,3 cures mice with SB-HCC mediated by CD8 T cells and associated with Th1 and Th17 responses.
From: Expression of tumor antigens within an oncolytic virus enhances the anti-tumor T cell response
A cDNA from three murine Sleeping Beauty HCC tumor-bearing livers, (SB HCC 1,2&3 Library), was pooled, cloned and amplified by PCR. The Library was then cloned into the VSV backbone plasmid between the Xho1 and Nhe1 sites. Figure created with BioRender.com under CC BY-NC-ND. B Composition of the VSV-derived ASMEL36 and SB HCC 1,2&3 Libraries was validated using PCR from the equivalent of 108 pfu of the VSV-ASMEL36 or VSV-SB HCC 1,2&3 with gene specific primers to the HCC-specific Lcn2, Lect2, and Smagp genes or to the melanoma specific TYRP1 and GP100 genes. All p values noted are <0.0001. C Following hydrodynamic injection of hMet + S45Y ß-Catenin (day 0), animals were treated on days 21, 23, 25, 28, 30, 32 with anti-PD-L1 (100 µg/injection) with, or without, 107 pfu of VSV-SB-HCC1,2,3 or 3 × 106 pfu each of (VSV-IFNß-Lect2 + VSV-IFNß-Lcn2 + VSV-IFNß-Smagp) on days 38,40,42. (N = 32, 8 mice per group). D, E 106 splenocytes were co-cultured with a 1:1;1 mixture of live SB-HCC 1,2,3 explant cells as targets at an effector:target ratio of 10:1 with supernatant assayed 48 h later for (D) IL-17 and (E) IFNγ. (N = 12, 3 mice per group, for (D) all p values noted are <0.0001, for (E) αPDL1 + VSV-Lcn2, -Lect2, -SMAGP vs. VSV-SB-HCC1,2,3 + αPDL1 p = 0.0230, VSV-SB-HCC1,2,3 + αPDL1 vs. VSV-SB-HCC1,2,3 + IgG p = 0.0437) (F). Following hydrodynamic injection of hMet + S45Y ß-Catenin (day 0), animals were treated on days 21, 23, 25, 28, 30, 32 with control isotype IgG or with anti-PD-L1 (200 µg/injection) with, or without, PBS or 107pfu ASMEL (VSV-melanoma cDNA library)36 or 107 pfu VSV-SB-HCC1,2,3 on days 38,40,42. Depleting antibodies were given concurrently with either anti-PD-L1 or IgG isotype control for 6 doses. (N = 65, 8 mice per group except VSV-SB-HCC1,2,3 + Control IgG, n = 9). Significance comparisons for (Fig. S8) are included in the Supplement, (B) was determined using 2-way ANOVA with multiple comparisons using Tukey’s multiple comparisons test, (C, F) were determined through survival curve comparison testing using a log-rank Mantel-Cox test, and (D, E) were determined through ordinary one-way ANOVA with Tukey’s multiple comparison tests.
