Summary of anti-hFIX IgG antibody formation in HB mice
Treatment group . | . | No. with antibodies* . | . | Average anti-hFIX IgG, mg/mL (range)† . | Average anti-hFIX antibody titer (range)‡ . | Average inhibitor titer, BU/mL (range)§ . | ||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Dose of RV . | hFIX protein injections . | ELISA . | Bethesda assay . | . | . | . | ||||||
| hFIX protein injections into HB mice that did not receive gene transfer∥ | ||||||||||||
| None | 8-10 without adjuvant (HB × 10) | 11 of 15 | 9 of 15 | 1.2 ± 0.3 (0.05-3.2) | 20 436 ± 6 140 (800-51 200) | 7.5 ± 2.2 (1.6-18) | ||||||
| None | 2 with adjuvant (HB × 2) | 10 of 10 | 10 of 10 | 2.1 ± 0.5 (0.6-4.6) | 143 000 ± 34 800 (40 000-320 000) | 9.1 ± 2.2 (1-18) | ||||||
| Neonatal gene transfer to HB mice before hFIX protein injections¶ | ||||||||||||
| High-dose RV, 1 × 1010 TU/kg | 10 without adjuvant | 0 of 6 P = .004 vs HB × 10 | 0 of 6 | 0 | < 1:100 | < 1 | ||||||
| Medium-dose RV, 1 × 109 TU/kg | 10 without adjuvant | 0 of 6 P = .004 vs HB × 10 | 0 of 6 | 0 | < 1:100 | < 1 | ||||||
| Low-dose RV, 1 × 108 TU/kg | None | 0 of 10 | 0 of 10 | 0 | < 1:100 | < 1 | ||||||
| 10 without adjuvant | 0 of 7 P = .004 vs HB × 10 | 0 of 7 | 0 | < 1:100 | < 1 | |||||||
| 10 without adjuvant and 1-2 with adjuvant | 1 of 4 P = .01 vs HB × 2 | 0 of 4 | 0.4 | 3 200 | < 1 | |||||||
| Very-low-dose RV, 1 × 107 TU/kg | None | 2 of 24 | 0 of 24 | 0.009 ± 0.003 (0.006 and 0.012) | 300 ± 100 (200 and 400) | < 1 | ||||||
| 10 without adjuvant | 1 of 3 NS vs HB × 10 | 0 of 3 | 0.21 | 6 400 | < 1 | |||||||
| 10 without adjuvant and 1-2 with adjuvant | 2 of 2 NS vs HB × 2 | 1 of 2 | 0.355 ± 0.345 (0.01-0.7) | 3 400 ± 3 000 (400 and 6 400) | 18 | |||||||
| 2 with adjuvant | 17 of 17 NS vs HB × 2 | 17 of 17 | 1.25 ± 0.4 (0.06-5.8) | 61 412 ± 18 017 (2 000-200 000) | 21.9 ± 10.3 (1.8-180) | |||||||
Treatment group . | . | No. with antibodies* . | . | Average anti-hFIX IgG, mg/mL (range)† . | Average anti-hFIX antibody titer (range)‡ . | Average inhibitor titer, BU/mL (range)§ . | ||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Dose of RV . | hFIX protein injections . | ELISA . | Bethesda assay . | . | . | . | ||||||
| hFIX protein injections into HB mice that did not receive gene transfer∥ | ||||||||||||
| None | 8-10 without adjuvant (HB × 10) | 11 of 15 | 9 of 15 | 1.2 ± 0.3 (0.05-3.2) | 20 436 ± 6 140 (800-51 200) | 7.5 ± 2.2 (1.6-18) | ||||||
| None | 2 with adjuvant (HB × 2) | 10 of 10 | 10 of 10 | 2.1 ± 0.5 (0.6-4.6) | 143 000 ± 34 800 (40 000-320 000) | 9.1 ± 2.2 (1-18) | ||||||
| Neonatal gene transfer to HB mice before hFIX protein injections¶ | ||||||||||||
| High-dose RV, 1 × 1010 TU/kg | 10 without adjuvant | 0 of 6 P = .004 vs HB × 10 | 0 of 6 | 0 | < 1:100 | < 1 | ||||||
| Medium-dose RV, 1 × 109 TU/kg | 10 without adjuvant | 0 of 6 P = .004 vs HB × 10 | 0 of 6 | 0 | < 1:100 | < 1 | ||||||
| Low-dose RV, 1 × 108 TU/kg | None | 0 of 10 | 0 of 10 | 0 | < 1:100 | < 1 | ||||||
| 10 without adjuvant | 0 of 7 P = .004 vs HB × 10 | 0 of 7 | 0 | < 1:100 | < 1 | |||||||
| 10 without adjuvant and 1-2 with adjuvant | 1 of 4 P = .01 vs HB × 2 | 0 of 4 | 0.4 | 3 200 | < 1 | |||||||
| Very-low-dose RV, 1 × 107 TU/kg | None | 2 of 24 | 0 of 24 | 0.009 ± 0.003 (0.006 and 0.012) | 300 ± 100 (200 and 400) | < 1 | ||||||
| 10 without adjuvant | 1 of 3 NS vs HB × 10 | 0 of 3 | 0.21 | 6 400 | < 1 | |||||||
| 10 without adjuvant and 1-2 with adjuvant | 2 of 2 NS vs HB × 2 | 1 of 2 | 0.355 ± 0.345 (0.01-0.7) | 3 400 ± 3 000 (400 and 6 400) | 18 | |||||||
| 2 with adjuvant | 17 of 17 NS vs HB × 2 | 17 of 17 | 1.25 ± 0.4 (0.06-5.8) | 61 412 ± 18 017 (2 000-200 000) | 21.9 ± 10.3 (1.8-180) | |||||||
Number of animals with significant anti-hFIX IgG antibodies was determined out of the total number of animals evaluated. P values were obtained by comparing the frequency of antibody formation using Fisher exact test for animals that received gene transfer with that in HB mice that received 10 injections of hFIX without adjuvant (HB × 10) or 2 injections of hFIX with adjuvant (HB × 2).
Average relative levels of anti-hFIX IgG ± SEM were determined using the highest value obtained for each animal with detectable antibodies.
Average anti-hFIX IgG titer was determined using the highest value for each animal that was positive.
Bethesda titer was determined for the sample with the highest anti-hFIX IgG antibody level and was averaged for all the animals that were positive.
Results are given for HB mice that did not receive gene transfer and were treated with 10 injections of 30 IU/kg per dose hFIX without adjuvant (HB × 10) or 2 doses of 30 IU/kg hFIX with adjuvant (HB × 2). These are the same mice whose antibody levels are shown in Figure 3B-C, respectively.
Results are given for HB mice that were injected with different doses of hAAT-hFIX-WPRE at birth. These are the same mice whose expression and antibody levels are shown in Figure 3A, D-H, respectively.