High levels of HbF in erythroblasts derived from PB CD34+ cells transduced with γ-globin lentiviral vectors
| Vector . | Cell proliferation* . | CD235+,† % . | GFP+,‡ % . | CFU-C, % vector+§ . | Vector copy no.‖ . | HbF,¶ % . |
|---|---|---|---|---|---|---|
| Mock | 250 ± 126 | 87 ± 11 | ND | ND | ND | 1.4 ± 0.4 |
| GFP | 230 ± 94 | 91 ± 9 | 87 ± 6 | ND | 3.5 ± 1.6 | 1.9 ± 0.5 |
| V5m3 | 319 ± 149 | 93 ± 7 | ND | 77 ± 3 | 0.9 ± 0.2 | 11.1 ± 3.1 |
| V5m3–400 | 259 ± 112 | 92 ± 5 | ND | 91 ± 9 | 1.0 ± 0.1 | 15.4 ± 4.9 |
| Vector . | Cell proliferation* . | CD235+,† % . | GFP+,‡ % . | CFU-C, % vector+§ . | Vector copy no.‖ . | HbF,¶ % . |
|---|---|---|---|---|---|---|
| Mock | 250 ± 126 | 87 ± 11 | ND | ND | ND | 1.4 ± 0.4 |
| GFP | 230 ± 94 | 91 ± 9 | 87 ± 6 | ND | 3.5 ± 1.6 | 1.9 ± 0.5 |
| V5m3 | 319 ± 149 | 93 ± 7 | ND | 77 ± 3 | 0.9 ± 0.2 | 11.1 ± 3.1 |
| V5m3–400 | 259 ± 112 | 92 ± 5 | ND | 91 ± 9 | 1.0 ± 0.1 | 15.4 ± 4.9 |
Data are mean ± SD.
ND indicates not done.
Fold increase in viable cells from day of transduction to culture day 11 or 12.
Erythroid differentiation was monitored by flow cytometric analysis of bulk cell populations for expression of CD235 (Glycophorin A) on culture day 14.
GFP+ fraction of bulk cell population determined by flow cytometry on culture days 12 to 14.
PCR analysis for the presence of vector sequences in clonogenic progenitors grown in methylcellulose supplemented with cytokines.
Genomic DNA was isolated from the bulk cell population on day 8 of culture and copy number determined by quantitative PCR or Southern blot analysis.
Percentage of fetal hemoglobin production quantified by HPLC and normalized per vector copy for V5m3 and V5m3–400 transduced cell populations.