AAV vector genome copy numbers per diploid liver cell genome and hFIX levels
. | Southern blot . | . | Q-PCR . | . | hFIX, μg/mL . | . | . | |||
|---|---|---|---|---|---|---|---|---|---|---|
| Vector . | Mean ± SD . | pv/tv . | Mean ± SD . | pv/tv . | Mean ± SD . | pv/tv . | hFIX/genome . | |||
| AAV-1 pv | 1.25 ± 0.6 | 11.36 | 0.725 ± 1.042 | 8.15 | 12.25 ± 6.16 | 9.49 | 16.9 | |||
| AAV-1 tv | 0.11 ± 0.06 | — | 0.089 ± 0.078 | — | 1.29 ± 0.78 | — | 14.49 | |||
| AAV-2 pv | 1.96 ± 1.02 | 28 | 0.52 ± 0.503 | 9.12 | 10.76 ± 3.33 | 7.63 | 20.69 | |||
| AAV-2 tv | 0.07 ± 0.01 | — | 0.057 ± 0.023 | — | 1.41 ± 0.75 | — | 24.74 | |||
| AAV-3 pv | 0.38 ± 0.15 | — | 0.097 ± 0.019 | 48.5 | 4.68 ± 1.27 | 78 | 48.25 | |||
| AAV-3 tv | < 0.03* | — | 0.002 ± 0.001 | — | 0.06 ± 0.03 | — | 30 | |||
| AAV-4 pv | < 0.03* | — | 0.001 | 1 | ND | — | — | |||
| AAV-4 tv | < 0.03* | — | 0.001 ± 0.001 | — | ND | — | — | |||
| AAV-5 pv | 0.03 ± 0.01 | — | 0.044 ± 0.041 | 14.67 | 0.93 ± 0.59 | — | 21.14 | |||
| AAV-5 tv | < 0.03* | — | 0.003 ± 0.001 | — | ND | — | — | |||
| AAV-6 pv | 1.61 ± 0.12 | 11.5 | 1 ± 0.587 | 8.93 | 18.7 ± 3.4 | 10.11 | 18.7 | |||
| AAV-6 tv | 0.14 ± 0.06 | — | 0.112 ± 0.036 | — | 1.85 ± 1.33 | — | 16.52 | |||
. | Southern blot . | . | Q-PCR . | . | hFIX, μg/mL . | . | . | |||
|---|---|---|---|---|---|---|---|---|---|---|
| Vector . | Mean ± SD . | pv/tv . | Mean ± SD . | pv/tv . | Mean ± SD . | pv/tv . | hFIX/genome . | |||
| AAV-1 pv | 1.25 ± 0.6 | 11.36 | 0.725 ± 1.042 | 8.15 | 12.25 ± 6.16 | 9.49 | 16.9 | |||
| AAV-1 tv | 0.11 ± 0.06 | — | 0.089 ± 0.078 | — | 1.29 ± 0.78 | — | 14.49 | |||
| AAV-2 pv | 1.96 ± 1.02 | 28 | 0.52 ± 0.503 | 9.12 | 10.76 ± 3.33 | 7.63 | 20.69 | |||
| AAV-2 tv | 0.07 ± 0.01 | — | 0.057 ± 0.023 | — | 1.41 ± 0.75 | — | 24.74 | |||
| AAV-3 pv | 0.38 ± 0.15 | — | 0.097 ± 0.019 | 48.5 | 4.68 ± 1.27 | 78 | 48.25 | |||
| AAV-3 tv | < 0.03* | — | 0.002 ± 0.001 | — | 0.06 ± 0.03 | — | 30 | |||
| AAV-4 pv | < 0.03* | — | 0.001 | 1 | ND | — | — | |||
| AAV-4 tv | < 0.03* | — | 0.001 ± 0.001 | — | ND | — | — | |||
| AAV-5 pv | 0.03 ± 0.01 | — | 0.044 ± 0.041 | 14.67 | 0.93 ± 0.59 | — | 21.14 | |||
| AAV-5 tv | < 0.03* | — | 0.003 ± 0.001 | — | ND | — | — | |||
| AAV-6 pv | 1.61 ± 0.12 | 11.5 | 1 ± 0.587 | 8.93 | 18.7 ± 3.4 | 10.11 | 18.7 | |||
| AAV-6 tv | 0.14 ± 0.06 | — | 0.112 ± 0.036 | — | 1.85 ± 1.33 | — | 16.52 | |||
AAV genomes were detected in livers from vector-treated animals and were quantified using Southern blot analysis or Q-PCR, as described in “Materials and methods.” Shown are absolute (Southern blot) or relative (Q-PCR) genome copy numbers per cell, together with hFIX levels expressed (mean ± SD; n = 3 per group). Within the Q-PCR data, values obtained from AAV-6 portal-vein-injected animals served as an internal reference and were set to 1 because of the lack of an appropriate absolute standard (see “Materials and methods”). Also shown are ratios of copy numbers of hFIX levels after portal or tail vein delivery of each vector (pv/tv). The rightmost column lists ratios of hFIX levels to relative genome copy numbers.
pv indicates portal vein; tv, tail vein;—, not applicable; and ND, not detected.
Below detection limit of the method.