Figure 4

(A) Experimental timeline of pooled human IgG, IdeZ, and AAV9-Luc injections. Cardiac tissues were derived as outlined earlier in the liver experiment. (B and D) Luciferase transgene expression levels were analyzed 4 weeks postinjection in the heart. Luciferase expression levels were normalized for total tissue protein concentration and represented as log relative luminescence units per gram of tissue. Each dot represents the average of a technical duplicate from a single animal. (C and E) Biodistribution of AAV9-Luc vector genomes in the heart. Vector genome copy numbers per cell were calculated based on normalization to copies of the lamin B2 housekeeping gene. Each dot represents the average of a technical duplicate from a single animal. Significance was determined by 1-way ANOVA with Tukey’s posttest. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (F) Schematic demonstrating experimental timeline of human serum, IdeZ, and AAV9-Luc injections. Cardiac tissues were derived as outlined earlier in the liver experiment. (G) Luciferase transgene expression levels were analyzed 4 weeks postinjection in the hearts of passively immunized mice treated with PBS (black bar, black circle) or prophylactically with IdeZ (gray bar, black triangle). Transduction levels were normalized to nonimmunized mice that were injected with AAV9-Luc at the same dose and represented as percentage of control. Each bar represents the average of a technical duplicate from a single animal. (H) Relative cardiac transduction efficiency of AAV9-Luc in the entire cohort of mice immunized with human sera treated with PBS control (circle) or IdeZ (triangle). Biodistribution of AAV9 vector genomes in the heart for mice passively immunized with individual human serum samples (I) and the entire cohort (J). Vector genome copy numbers per cell were calculated based on normalization to copies of the lamin B2 housekeeping gene. Each bar represents the average of a technical duplicate from a single animal. Significance was determined by the nonparametric Mann-Whitney rank test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.