Fig. 5: Assessment of immunogenicity of AdVLP-7 by microneutralization assay.

Neutralizing antibody titers in sera collected from AdVLP-7-immunized mice, WT AdV-7-immunized mice, and mice injected with a sham (n = 10/group) were measured using a microneutralization assay based on a recombinant AdV-7 that expresses GFP (rAdV-7). a Representative images of A549 cells taken 28 h after infection with rAdV-7 that had been mixed with serum from a sample animal in each group at a 1/250 dilution. Scale bars are 200 µm. b Neutralizing antibody titers, presented as the serum dilution at which half of the rAdV-7 was neutralized as compared to serum-negative controls (ID₅₀). ID₅₀ values for individual animals are presented as either circles (•) for females or triangles (▼) for males. Dotted line indicates lower limit of detection (starting serum dilution). In (b), data are mean + SD. Results were analyzed by one-way ANOVA with Tukey’s multiple comparisons test, with p < 0.05 considered statistically significant. ^****p < 0.0001. All treatment groups were significant as compared to Sham (p < 0.0001). c–f Correlation analysis between neutralizing antibody titers (ID₅₀) and binding antibody titers against (c) AdV-7, (d) hexon, (e) penton, or (f) fiber (as determined by ELISA, see Figs. 3–4). In (c–f), data were analyzed by two-tailed Pearson’s correlation analysis, alpha = 0.05. R² and p-values are presented for each correlation.