Transduction rate of HATGF-βRII-Δcyt ranges from 6.5% to 55% as determined by quantitative real-time polymerase chain reaction
| . | No. of transduced copies per 100 000 cells . | % transduced* . |
|---|---|---|
| CTL-sorted (positive selection of transduced cells by FACS) | 310 000 | 100 |
| Donor 1 (unsorted) | 67 550 | 19 |
| Donor 2 (unsorted) | 28 055 | 10 |
| Patient 1 (unsorted) | 141 000 | 45 |
| Patient 3 (unsorted) | 170 000 | 55 |
| Patient 4 (unsorted) | 22 140 | 6.5 |
| . | No. of transduced copies per 100 000 cells . | % transduced* . |
|---|---|---|
| CTL-sorted (positive selection of transduced cells by FACS) | 310 000 | 100 |
| Donor 1 (unsorted) | 67 550 | 19 |
| Donor 2 (unsorted) | 28 055 | 10 |
| Patient 1 (unsorted) | 141 000 | 45 |
| Patient 3 (unsorted) | 170 000 | 55 |
| Patient 4 (unsorted) | 22 140 | 6.5 |
To assess the transduction efficiency, DNA was extracted from transduced cytotoxic T lymphocytes (CTLs) and transduced CTLs sorted for the hemagglutinin peptide epitope tag. These sorted cells were therefore assumed to be 100% transduced. The number of copies per 100 000 cells was calculated by using a standard curve generated by serial dilutions of plasmid DNA from 256 to 131 072 copies. Sorted CTLs were compared with 5 unsorted transduced CTL lines, and the transduction rates were calculated.
Compared with 100% population of sorted CTLs.