TABLE 1.
Hepatocyte transduction with AAV1-, AAV2-, or AAV8-EF1α-nlslacZ 6 weeks postinjection
| Vector | Dose (vg/mouse) | No. of mice | Mean transduction efficiency (%)a ± SD | Mean β-galactosidase expression (pg/mg of protein)b ± SD | Mean vector copy no. (ds-vg/dge) ± SD | Mean net vector copy no. per transduced cell (ds-vg/dge)c ± SD | Mean β-galactosidase expression rated ± SD | Mean vector genome activitye ± SD |
|---|---|---|---|---|---|---|---|---|
| AAV1 | 5.0 × 1010 | 4 | 4.2 ± 1.0 | 426 ± 110 | 1.1 ± 0.2 | 27.0 ± 3.1 | 100.5 ± 9.5 | 376.7 ± 67.4 |
| 3.0 × 1011 | 6 | 7.7 ± 0.7 | 1215 ± 263 | 9.3 ± 0.9 | 121.6 ± 14.2 | 160.1 ± 40.5 | 133.3 ± 38.4 | |
| 1.8 × 1012 | 4 | 13.2 ± 2.5 | 2078 ± 389 | 54.9 ± 18.1 | 413.5 ± 83.1 | 159.7 ± 27.8 | 40.1 ± 12.1 | |
| 7.2 × 1012 | 3 | 23.5 ± 1.0 | 2057 ± 99 | 92.3 ± 12.2 | 412.1 ± 69.6 | 88.9 ± 8.0 | 21.5 ± 1.6 | |
| AAV2 | 3.0 × 1011 | 6 | 3.9 ± 0.5 | 888 ± 228 | 5.1 ± 2.0 | 132.2 ± 47.5 | 235.8 ± 72.4 | 197.0 ± 92.5 |
| AAV8 | 5.0 × 1010 | 4 | 8.1 ± 1.8 | 1669 ± 201 | 7.2 ± 1.5 | 90.6 ± 5.3 | 211.7 ± 31.5 | 223.5 ± 23.5 |
| 3.0 × 1011 | 6 | 14.9 ± 3.4 | 2963 ± 517 | 58.2 ± 10.9 | 413.1 ± 134.8 | 208.2 ± 63.5 | 53.0 ± 14.6 | |
| 1.8 × 1012 | 3 | 65.8 ± 9.0 | 9052 ± 1541 | 207.5 ± 56.0 | 318.4 ± 87.5 | 140.8 ± 40.2 | 44.8 ± 7.4 | |
| 7.2 × 1012 | 4 | 97.4 ± 0.3 | 21686 ± 3051 | 620.6 ± 42.8 | 637.2 ± 42.6 | 222.7 ± 31.4 | 35.3 ± 7.2 | |
| None (excipient) | 3 | 0.0 | <4 | 0.0 | 0.0 | NA | NA |
X-Gal-positive nuclei/total hepatocyte nuclei counted (at least 2,000 hepatocyte nuclei were counted).
β-Galactosidase antigen levels in liver extracts were normalized to the amount of total protein in samples.
Net vector copy number is defined as the number of double-stranded vector genomes per transduced hepatocyte; i.e., (vector copy number per cell [vg/dge]/transduction efficiency [%]) × 100.
β-Galactosidase expression rate is defined as picograms of β-galactosidase protein per milligram of cellular protein produced from transduced hepatocytes corresponding to 1% of total hepatocytes; i.e., β-galactosidase expression divided by transduction efficiency. The values reflect β-galactosidase production per transduced hepatocyte. NA, not applicable.
Vector genome activity is defined as β-galactosidase production per double-stranded vector genome; i.e., β-galactosidase expression divided by vector copy number (ds-vg/dge).