MDR1 provirus–marked human cells detected in BM of chimeric mice by quantitative real-time PCR and by end point PCR on single colonies
| Transduction Group . | NOD/SCID Set No. . | MDR1 Provirus-Marked Human Cells in BM (%) . | MDR1-PCR+ Human ColonyForming Cells (%) . |
|---|---|---|---|
| Transwell | |||
| IL-3 | 3, 4 | NA | No DNA |
| IL-3 | 5, 7 | 4 ± 2 | NA |
| IL-3 + SCM | 1, 7 | 5 ± 1 | 5 |
| IL-3/FL | 3, 4 | NA | No DNA |
| IL-3/FL | 5, 6, 7, 8 | 3 ± 1 | NA |
| IL-3/FL + SCM | 1, 2, 7 | 20 | 2 ± 2 |
| IL-3/IL-6/SCF/FL | 8 | 12 ± 2 | NA |
| FL/T/SCF | 8 | 17 ± 7 | 10 |
| FL/T/V | 7 | 1 ± 1 | NA |
| Cell-free viral supernatant | |||
| IL-3/IL-6/SCF/FL | 10 | 11 ± 3 | 23 ± 3 |
| FL/T/SCF | 10 | 38 ± 12 | 32 ± 9 |
| FL/T/SCF | 9 | 20 ± 6 | 16 ± 6 |
| Transduction Group . | NOD/SCID Set No. . | MDR1 Provirus-Marked Human Cells in BM (%) . | MDR1-PCR+ Human ColonyForming Cells (%) . |
|---|---|---|---|
| Transwell | |||
| IL-3 | 3, 4 | NA | No DNA |
| IL-3 | 5, 7 | 4 ± 2 | NA |
| IL-3 + SCM | 1, 7 | 5 ± 1 | 5 |
| IL-3/FL | 3, 4 | NA | No DNA |
| IL-3/FL | 5, 6, 7, 8 | 3 ± 1 | NA |
| IL-3/FL + SCM | 1, 2, 7 | 20 | 2 ± 2 |
| IL-3/IL-6/SCF/FL | 8 | 12 ± 2 | NA |
| FL/T/SCF | 8 | 17 ± 7 | 10 |
| FL/T/V | 7 | 1 ± 1 | NA |
| Cell-free viral supernatant | |||
| IL-3/IL-6/SCF/FL | 10 | 11 ± 3 | 23 ± 3 |
| FL/T/SCF | 10 | 38 ± 12 | 32 ± 9 |
| FL/T/SCF | 9 | 20 ± 6 | 16 ± 6 |
Real-time PCR was performed on genomic BM DNA to determine the percentage of provirus-marked human cells. In all experiments, genomic DNA from NOD/SCID mice not given transplanted cells and from control mice that had mock-transduced CD34+ PBPC transplantation was concurrently analyzed to verify the specificity of the PCR reactions for the provirus. BM cells from chimeric mice were plated in semisolid colony assays specific for human progenitors. Individual human-cell colonies derived from cultures under nonselective conditions were plucked and subjected to a nested PCR analysis of SF-MDR proviral DNA.
Values are mean ± SE unless otherwise indicated.