TABLE 3.
Hepatocyte transduction with AAV-EF1α-nlsLacZ (28 weeks postinjection)
| Dose (vg/mouse) | Mouse | Transduction efficiency (%)a | β-Gal expression (pg/mg of protein)b | Vector copy no. (vg/dge)
|
Avg (mean ± SD)
|
||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Pre-PHxc | Post-PHx | Pre-PHx
|
Post-PHx copy no. % dropg | ||||||||
| Transduction efficiency (%) | β-Gal expression (pg/mg of protein) | Vector copy no. (vg/dge) | β-Gal expression rated | Net vector copy no./transduced celle (vg/dge) | |||||||
| 5.0 × 1010 | 1 | 0.6 | 88 | 0.13 | 0.01 | 0.6 ± 0.4 | 93 ± 55 | 0.19 ± 0.08 | 183 ± 137 | 40 ± 18 | 89 ± 4 |
| 2 | 0.3 | 20 | 0.15 | 0.02 | |||||||
| 3 | 0.3 | 115 | 0.18 | ||||||||
| 4 | 1.1 | 150 | 0.30 | 0.02 | |||||||
| 3.0 × 1011 (6×)f | 5 | 2.3 | 574 | 3.91 | 0.39 | 3.0 ± 0.5 (5.0×) | 544 ± 333 (5.8×) | 2.14 ± 1.26 (11.3×) | 187 ± 106 | 79 ± 62 | 77 ± 11 |
| 6 | 3.0 | 394 | 0.94 | 0.43 | |||||||
| 7 | 3.2 | 217 | 1.85 | 0.46 | |||||||
| 8 | 3.3 | 922 | 1.84 | 0.24 | |||||||
| 1.8 × 1012 (6×) | 9 | 7.8 | 2,152 | 13.61 | 8.1 ± 1.0 (2.7×) | 1,552 ± 676 (2.9×) | 15.57 ± 2.83 (7.4×) | 192 ± 88 | 196 ± 60 | 88 ± 3 | |
| 10 | 9.1 | 1,529 | 13.46 | 2.10 | |||||||
| 11 | 8.7 | 1,138 | 15.66 | 1.72 | |||||||
| 12 | 6.9 | 1,026 | 19.54 | 1.92 | |||||||
| 3.9 × 1012 (2.2×) | 13 | 8.9 | 1,237 | 16.22 | 1.50 | 8.9 (1.1×) | 1,237 (0.8×) | 16.22 (1.0×) | 138 | 182 | 91 |
X-Gal-positive nuclei/total hepatocyte nuclei counted (at least 2,000 hepatocyte nuclei were counted).
β-Gal antigen levels in liver extracts were normalized by total protein in samples.
PHx, partial hepatectomy.
β-Gal expression rate is defined as an amount of β-Gal protein per milligram of cellular protein (picograms per milligram of protein) produced from transduced hepatocytes corresponding to 1% of total hepatocytes, i.e., β-Gal expression (picograms per milligram of protein) divided by transduction efficiency (percent). The values reflect β-Gal protein production per transduced hepatocyte.
Net vector copy number is defined as the number of vector genomes per transduced hepatocyte, i.e., (vector copy number per cell [vg/dge]/transduction efficiency [%]) × 100.
Numbers in parentheses indicate increments or increases compared to 1 lower dose.
Levels of β-Gal expression after partial hepatectomy were not evaluated because of an unexpectedly long half-life of this protein in mouse hepatocytes (Nakai et al., unpublished).