Summary of anti-hFIX IgG antibody formation in normal mice
Treatment group and mouse strain . | No. with antibodies* . | . | Average IgG, relative mg/mL (range)† . | . | Average inhibitor titer, BU/mL (range)§ . | |||||
|---|---|---|---|---|---|---|---|---|---|---|
| . | ELISA . | Bethesda assay . | . | Average ELISA titer (range)‡ . | . | |||||
| Protein injections without gene transfer∥ | ||||||||||
| C3H | 7 of 7 | 7 of 7 | 3.8 ± 1.4 (0.5-10.6) | 263 314 ± 132 467 (102 400-409 600) | 16 ± 2 (6-20) | |||||
| BALB/c: 129S | 8 of 8 | 8 of 8 | 12.2 ± 5.2 (0.6-34.4) | 421 410 ± 169 746 (12 800-1 000 000) | 80 ± 30 (12-200) | |||||
| BALB/c | 6 of 8 | 6 of 8 | 13.8 ± 4.6 (0.2-25.9) | 1 097 600 ± 342 040 (6 400-1 638 400) | 92 ± 32 (1.8-180) | |||||
| C57BL/6 | 5 of 8 | 2 of 8 | 1.7 ± 1.3 (0.04-6.7) | 24 000 ± 19 622 (1 600-102 400) | 9 ± 8 (1 and 18) | |||||
| High-dose neonatal gene transfer followed by 10 hFIX injections without adjuvant and 2 hFIX injections with adjuvant¶ | ||||||||||
| C3H | 0 of 5 P = .0008 | 0 of 5 | 0 | < 1:100 | < 1 | |||||
| BALB/c: 129S | 0 of 3 P = .006 | 0 of 3 | 0 | < 1:100 | < 1 | |||||
| BALB/c | 0 of 7 P = .007 | 0 of 7 | 0 | < 1:100 | < 1 | |||||
| High-dose neonatal gene transfer without protein injections# | ||||||||||
| C57BL/6 | 0 of 3 NS | 0 of 3 | 0 | < 1:100 | < 1 | |||||
Treatment group and mouse strain . | No. with antibodies* . | . | Average IgG, relative mg/mL (range)† . | . | Average inhibitor titer, BU/mL (range)§ . | |||||
|---|---|---|---|---|---|---|---|---|---|---|
| . | ELISA . | Bethesda assay . | . | Average ELISA titer (range)‡ . | . | |||||
| Protein injections without gene transfer∥ | ||||||||||
| C3H | 7 of 7 | 7 of 7 | 3.8 ± 1.4 (0.5-10.6) | 263 314 ± 132 467 (102 400-409 600) | 16 ± 2 (6-20) | |||||
| BALB/c: 129S | 8 of 8 | 8 of 8 | 12.2 ± 5.2 (0.6-34.4) | 421 410 ± 169 746 (12 800-1 000 000) | 80 ± 30 (12-200) | |||||
| BALB/c | 6 of 8 | 6 of 8 | 13.8 ± 4.6 (0.2-25.9) | 1 097 600 ± 342 040 (6 400-1 638 400) | 92 ± 32 (1.8-180) | |||||
| C57BL/6 | 5 of 8 | 2 of 8 | 1.7 ± 1.3 (0.04-6.7) | 24 000 ± 19 622 (1 600-102 400) | 9 ± 8 (1 and 18) | |||||
| High-dose neonatal gene transfer followed by 10 hFIX injections without adjuvant and 2 hFIX injections with adjuvant¶ | ||||||||||
| C3H | 0 of 5 P = .0008 | 0 of 5 | 0 | < 1:100 | < 1 | |||||
| BALB/c: 129S | 0 of 3 P = .006 | 0 of 3 | 0 | < 1:100 | < 1 | |||||
| BALB/c | 0 of 7 P = .007 | 0 of 7 | 0 | < 1:100 | < 1 | |||||
| High-dose neonatal gene transfer without protein injections# | ||||||||||
| C57BL/6 | 0 of 3 NS | 0 of 3 | 0 | < 1:100 | < 1 | |||||
NS indicates not significant.
Number of animals with significant anti-hFIX IgG antibodies was determined from 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 mice of the same strain that did not receive gene transfer but received 10 injections of hFIX without adjuvant.
Average relative levels of anti-hFIX IgG ± SEM were determined using the highest value obtained for each animal with detectable antibodies.
Average titer was determined using the highest value for each animal that was positive.
Bethesda titer was determined for the sample with the highest levels of anti-hFIX IgG antibody in the immunoassay.
Results are given for animals of the indicated strain that did not receive gene transfer and were treated with 10 injections of 30 IU/kg per dose hFIX without adjuvant. These are the same mice whose antibody levels are shown in Figure 2A.
Results are given for mice of the indicated strain that were injected with 1 × 1010 TU/kg hAAT-hFIX-WPRE at birth and then received 10 injections of 30 IU/kg per dose hFIX without adjuvant followed by 2 injections of 30 IU/kg hFIX with adjuvant. These are the same mice whose expression and antibody levels are shown in Figures 1B and 2B, respectively.
Results are given for mice of the indicated strain that were injected with 1 × 1010 TU/kg hAAT-hFIX-WPRE at birth and were not challenged with protein. These are the same mice whose expression levels are shown in Figure 1B.