Threshold for Pre-existing Antibody Levels Limiting Transduction Efficiency of Systemic rAAV9 Gene Delivery: Relevance for Translation

Figures

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  Figure 1. Impact of Pre-existing αAAV9 Abs on the Biodistribution of Vector Genome in NHPs after an i.v. Injection of rAAV9 Vector Seven NHPs (age, 2–2.6 years) with different levels of pre-existing αAAV9 Abs were treated with an i.v. injection of 1 × 1013 vg/kg rAAV9-CBA-hNAGLUop vector. Necropsies were performed for tissue analyses at 1 month p.i. Tissues were assayed by qPCR for level of vector genome (vg). Data are expressed as 105 vg/μg gDNA. Ab-High: tissues from animals with 1:800–1:6,400 serum total αAAV9-IgG (n = 3); Ab-Low, tissues from animals with 1:100–1:400 serum total αAAV9-IgG (n = 4); Ab-Neg, tissues from αAAV9-IgG-negative NHPs from a previous study, treated with rAAV9-hNAGLU vector (n = 2).

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  Figure 2. Impact of Pre-existing αAAV9 Abs on Transgene Expression Levels in NHPs after an i.v. Injection of rAAV9 Vector Seven NHPs (age, 2–2.6 years) with different levels of pre-existing αAAV9 Abs were treated with an i.v. injection of 1 × 1013 vg/kg rAAV9-CBA-hNAGLUop vector. Necropsies were performed for tissue analyses at 1 month p.i. Tissues were assayed for NAGLU activity, expressed as U/mg protein: 1 U = 1 nmol 4MU released/h. H1–3, tissues from animals with 1:800–1:6,400 serum total αAAV9-IgG (n = 3); L1–4, tissues from animals with 1:100–1:400 serum total αAAV9-IgG (n = 4); NT, tissues from non-treated control animals from a previous study (n = 2).

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  Figure 3. Correlation of Tissue Vector Genome and NAGLU Activity in NHPs with Different Levels of Pre-existing αAAV9 Abs after an i.v. Injection of rAAV9 vector Seven NHPs (age, 2–2.6 years) with different levels of pre-existing αAAV9 Abs were treated with an i.v. injection of 1 × 1013 vg/kg rAAV9-CBA-hNAGLUop vector. Necropsies were performed for tissue analyses at 1 month p.i. Tissues were assayed for vector genome (105 vg/μg gDNA) and NAGLU activity (U/mg protein). Data were analyzed for correlation between tissue vg and NAGLU activity. Solid circles, tissues from animals with 1:800–1:6,400 serum total αAAV9-IgG (n = 3); empty circles, tissues from animals with 1:100–1:400 serum total αAAV9-IgG (n = 4).

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  Figure 4. Correlation between Vector Genome Biodistribution and Serum Levels of Pre-existing αAAV9 Abs in NHPs Treated with an i.v. Injection of rAAV9 Vector Seven NHPs (age 2–2.6 years) with different levels of pre-existing αAAV9 Abs were treated with an i.v. injection of 1 × 1013 vg/kg rAAV9-CBA-hNAGLUop vector. Necropsies were performed for tissue analyses at 1 month p.i. Data were analyzed for correlation between tissue vg (105 vg/μg gDNA) and serum levels of pre-existing αAAV9-Abs (ELISA titer). Solid circles, tissues from animals with 1:800–1:6,400 serum total αAAV9-IgG (n = 3); empty circles, tissues from animals with 1:100–1:400 serum total αAAV9-IgG (n = 4).

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  Figure 5. Correlation between Tissue NAGLU Activity and Serum Levels of Pre-existing αAAV9 Abs in NHPs Treated with an i.v. Injection of rAAV9 Vector Seven NHPs (age, 2–2.6 years) with different levels of pre-existing αAAV9 Abs were treated with an i.v. injection of 1 × 1013 vg/kg rAAV9-CBA-hNAGLUop vector. Necropsies were performed for tissue analyses at 1 month p.i. Data were analyzed for correlation between tissue NAGLU activity (U/mg) and serum levels of pre-existing αAAV9 Abs (ELISA titer). Solid circles, tissues from animals with 1:800–1:6,400 serum total αAAV9-IgG (n = 3); empty circles, tissues from animals with 1:100–1:400 serum total αAAV9-IgG (n = 4).