Summary of rhesus macaques treated with AAV8-hFIX vectors
Animal no. according to group . | Preexisting AAV8 NAB . | Vector dose, vg/kg . | Peak hFIX level,*ng/mL (% of normal) . | Steady-state hFIX level,*ng/mL (% of normal) mean ± SEM . | hFIX DNA in liver,†copy/diploid genome, mean ± SEM . |
|---|---|---|---|---|---|
| Efficacy study‡ | |||||
| R1 | < 1:1 | 5 × 1012 | < 39 (< 0.7) | < 39 (< 0.7) | 0.5 ± 0.2 |
| R2 | < 1:1 | 5 × 1012 | 390 (7.8) | 75 ± 6 (1.5 ± 0.1) | 3.3 ± 1.2 |
| R3 | < 1:1 | 5 × 1012 | 2074 (41.5) | 479 ± 61 (9.6 ± 1.2) | 8.5 ± 2.5 |
| R4 | < 1:1 | 5 × 1012 | 488 (9.8) | 208 ± 18 (4.2 ± 0.4) | 7.5 ± 2.3 |
| R5 | < 1:1 | 5 × 1012 | 538 (10.8) | 271 ± 29 (5.4 ± 0.6) | 5.1 ± 1.5 |
| R6 | < 1:1 | 5 × 1012 | 660 (13.2) | 312 ± 42 (6.2 ± 0.8) | 5.1 ± 1.8 |
| R7 | 1:5 | 5 × 1012 | < 39 (< 0.7) | < 39 (< 0.7) | 0.12 ± 0.01 |
| R8 | 1:5 | 5 × 1012 | < 39 (< 0.7) | < 39 (< 0.7) | 0.01 ± 0.03 |
| R9 | 1:5 | 5 × 1012 | < 39 (< 0.7) | < 39 (< 0.7) | 0.10 ± 0.05 |
| R10 | 1:5 | 5 × 1012 | < 39 (< 0.7) | < 39 (< 0.7) | 0.15 ± 0.01 |
| Immunosuppression study | |||||
| No IS regimen | |||||
| R11 | < 1:1 | 2 × 1013 | 1386 (27.7) | 206 ± 22 (4.1 ± 0.4) | ND |
| R12 | < 1:1 | 2 × 1013 | 697 (13.9) | 520 ± 48 (10.4 ± 1.0) | 1.1 ± 0.7 |
| R13 | < 1:1 | 2 × 1013 | 3396 (67.9) | 778 ± 147 (15.6 ± 2.9) | ND |
| IS regimen§ | |||||
| R14 | < 1:1 | 2 × 1013 | 2669 (53.4) | 920 ± 316 (18.4 ± 6.3) | 3.7 ± 2.4 |
| R15 | < 1:1 | 2 × 1013 | 480 (9.6) | 370 ± 30 (7.4 ± 0.6) | ND |
| R16 | < 1:1 | 2 × 1013 | 305 (6.1) | 150 ± 9 (3.0 ± 0.2) | ND |
Animal no. according to group . | Preexisting AAV8 NAB . | Vector dose, vg/kg . | Peak hFIX level,*ng/mL (% of normal) . | Steady-state hFIX level,*ng/mL (% of normal) mean ± SEM . | hFIX DNA in liver,†copy/diploid genome, mean ± SEM . |
|---|---|---|---|---|---|
| Efficacy study‡ | |||||
| R1 | < 1:1 | 5 × 1012 | < 39 (< 0.7) | < 39 (< 0.7) | 0.5 ± 0.2 |
| R2 | < 1:1 | 5 × 1012 | 390 (7.8) | 75 ± 6 (1.5 ± 0.1) | 3.3 ± 1.2 |
| R3 | < 1:1 | 5 × 1012 | 2074 (41.5) | 479 ± 61 (9.6 ± 1.2) | 8.5 ± 2.5 |
| R4 | < 1:1 | 5 × 1012 | 488 (9.8) | 208 ± 18 (4.2 ± 0.4) | 7.5 ± 2.3 |
| R5 | < 1:1 | 5 × 1012 | 538 (10.8) | 271 ± 29 (5.4 ± 0.6) | 5.1 ± 1.5 |
| R6 | < 1:1 | 5 × 1012 | 660 (13.2) | 312 ± 42 (6.2 ± 0.8) | 5.1 ± 1.8 |
| R7 | 1:5 | 5 × 1012 | < 39 (< 0.7) | < 39 (< 0.7) | 0.12 ± 0.01 |
| R8 | 1:5 | 5 × 1012 | < 39 (< 0.7) | < 39 (< 0.7) | 0.01 ± 0.03 |
| R9 | 1:5 | 5 × 1012 | < 39 (< 0.7) | < 39 (< 0.7) | 0.10 ± 0.05 |
| R10 | 1:5 | 5 × 1012 | < 39 (< 0.7) | < 39 (< 0.7) | 0.15 ± 0.01 |
| Immunosuppression study | |||||
| No IS regimen | |||||
| R11 | < 1:1 | 2 × 1013 | 1386 (27.7) | 206 ± 22 (4.1 ± 0.4) | ND |
| R12 | < 1:1 | 2 × 1013 | 697 (13.9) | 520 ± 48 (10.4 ± 1.0) | 1.1 ± 0.7 |
| R13 | < 1:1 | 2 × 1013 | 3396 (67.9) | 778 ± 147 (15.6 ± 2.9) | ND |
| IS regimen§ | |||||
| R14 | < 1:1 | 2 × 1013 | 2669 (53.4) | 920 ± 316 (18.4 ± 6.3) | 3.7 ± 2.4 |
| R15 | < 1:1 | 2 × 1013 | 480 (9.6) | 370 ± 30 (7.4 ± 0.6) | ND |
| R16 | < 1:1 | 2 × 1013 | 305 (6.1) | 150 ± 9 (3.0 ± 0.2) | ND |
ND indicates not determined.
Circulating hFIX levels in rhesus plasma were determined by ELISA (see “Materials and methods”), the limit of detection is 39 ng/mL.
Mean copy numbers of hFIX transgene DNA in liver as determined by Southern blotting of gDNAs from different hepatic lobes.
Additional 5 rhesus macaques which had anti-AAV8 titers ∼1:25 served as uninjected controls. The controls were housed in the same room and blood analyses and liver biopsies were performed along with the study subjects to monitor for any spontaneous variables. In all cases, the controls remained healthy with normal laboratory findings (data not shown).
Six-week oral treatment (twice/d) of immunosuppressants: tacrolimus and mycophenolate mofetil (see “Materials and methods”).