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(A) Experimental design. All mice received two doses of HSV-1 gD mRNA administered one week apart. Mice were then assigned to receive PBS, mRNA-UB-E6/E7-gD, ADU-S100, or ADU-S100 + mRNA-UB-E6/E7-gD. One week after the final vaccination, mice were challenged subcutaneously with TC-1 cells. ADU-S100 (10 μg/dose) was administered intratumorally beginning 7 days after tumor implantation, when tumors reached approximately 5 × 5 mm, for a total of three treatments. (B) Tumor volumes of individual mice at the experimental endpoint. (C) Kaplan–Meier survival analysis of the indicated treatment groups. (D) Cytotoxic T-lymphocyte (CTL) activity against TC-1 target cells measured 24 h after assay initiation. Effector-to-target (E) ratios are indicated. (E) IFN-γ production in splenocyte culture supernatants measured by ELISA 24 h after assay initiation. E ratios are indicated. (F, G) Flow cytometric analysis of HPV16 E7-specific CD8 + T cells in tumor-infiltrating lymphocytes (F) and splenocytes (G), determined by HPV16 E7 tetramer staining. (H) Quantification of CD4 + and CD8 + tumor-infiltrating cells by immunohistochemistry. Positive cells were quantified using ImageJ from randomly selected microscopic fields and expressed as the number of positive cells per field. Statistical significance: ns, not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

Journal: Research Square

Article Title: Stepwise Enhancement of HPV16 E6/E7 mRNA Vaccine Efficacy Using HSV-1 gD Epitope Incorporation and Immune-Modulating Agents

doi: 10.21203/rs.3.rs-9890379/v1

Figure Lengend Snippet: (A) Experimental design. All mice received two doses of HSV-1 gD mRNA administered one week apart. Mice were then assigned to receive PBS, mRNA-UB-E6/E7-gD, ADU-S100, or ADU-S100 + mRNA-UB-E6/E7-gD. One week after the final vaccination, mice were challenged subcutaneously with TC-1 cells. ADU-S100 (10 μg/dose) was administered intratumorally beginning 7 days after tumor implantation, when tumors reached approximately 5 × 5 mm, for a total of three treatments. (B) Tumor volumes of individual mice at the experimental endpoint. (C) Kaplan–Meier survival analysis of the indicated treatment groups. (D) Cytotoxic T-lymphocyte (CTL) activity against TC-1 target cells measured 24 h after assay initiation. Effector-to-target (E) ratios are indicated. (E) IFN-γ production in splenocyte culture supernatants measured by ELISA 24 h after assay initiation. E ratios are indicated. (F, G) Flow cytometric analysis of HPV16 E7-specific CD8 + T cells in tumor-infiltrating lymphocytes (F) and splenocytes (G), determined by HPV16 E7 tetramer staining. (H) Quantification of CD4 + and CD8 + tumor-infiltrating cells by immunohistochemistry. Positive cells were quantified using ImageJ from randomly selected microscopic fields and expressed as the number of positive cells per field. Statistical significance: ns, not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

Article Snippet: ADU-S100 [2′3′-c-di-AM(PS)2 (Rp,Rp); MedChemExpress; cat. no. HY-12885] was administered intratumorally at 10 μg per mouse for three total doses given every 3 days.

Techniques: Tumor Implantation, Activity Assay, Enzyme-linked Immunosorbent Assay, Staining, Immunohistochemistry