Transplantation of oncoretrovirally and lentivirally transduced cells
Animal . | Vector . | Pseudotype . | MOI . | Growth factors . | FACS+* . | Colonies/ 1000 cells plated† . | PCR+‡ . | Cells infused, × 106/kg . | Days to ANC > 500/μL . | Peak marking, % (day) . | Lost marking, days . | Follow-up, days . |
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| M00067 | RRLsin. CMVGFP (lenti) | VSV-G | 10 | 36SMFG | < 1 | 25 | 1 | 6.1 | 15 | 0.1 (11) | 54 | 180 |
| RRLsin.cPPT.PGKYFP.Wpre (lenti) | VSV-G | 10 | 36SMFG | 3 | 23 | 30 | 0.8 (11) | 54 | ||||
| A00083 | RRLsin.cPPT.PGKYFP.Wpre (lenti) | VSV-G | 100 | 36SMFG | 52 | ND | 80 | 8.2 | 23 | 3.6 (14) | 35 | 181 |
| RRLsin.cPPT.PGKGFP.Wpre (lenti) | VSV-G | 100 | S | 41 | ND | 36 | 0.4 (14) | 28 | ||||
| A00074 | RRLsin.cPPT.PGKYFP.Wpre (lenti) | VSV-G | 100 | 6SMF | 16 | ND | ND | 9.1 | 20 | 0.9 (9) | 26 | 177 |
| RRLsin.cPPT.PGKGFP.Wpre (lenti) | VSV-G | 10 | 6SMF | 6 | ND | ND | 0.1 (9) | 13 | ||||
| K99307 | RRLsin.cPPT.PGKGFP.Wpre (lenti) | VSV-G | 100 | 36SMFG | 17 | 26 | 55 | 19.7 | 20 | 1.1 (10) | 27 | 180 |
| MNDEYFPSN (onco) | Phoenix-GALV | 1 | 36SMFG | 50 | 26 | 70 | 23.2 (10) | 47 | ||||
| M99149 | RRLsin.cPPT.PGKGFP.Wpre (lenti) | VSV-G | 100 | 36SMFG | 6 | 33 | 40 | 17.6 | 20 | 0.3 (10) | 17 | 180 |
| MNDEYFPSN (onco) | Phoenix-GALV | 1 | 36SMFG | 34 | 29 | 55 | 12.1 (10) | 17 |
Animal . | Vector . | Pseudotype . | MOI . | Growth factors . | FACS+* . | Colonies/ 1000 cells plated† . | PCR+‡ . | Cells infused, × 106/kg . | Days to ANC > 500/μL . | Peak marking, % (day) . | Lost marking, days . | Follow-up, days . |
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| M00067 | RRLsin. CMVGFP (lenti) | VSV-G | 10 | 36SMFG | < 1 | 25 | 1 | 6.1 | 15 | 0.1 (11) | 54 | 180 |
| RRLsin.cPPT.PGKYFP.Wpre (lenti) | VSV-G | 10 | 36SMFG | 3 | 23 | 30 | 0.8 (11) | 54 | ||||
| A00083 | RRLsin.cPPT.PGKYFP.Wpre (lenti) | VSV-G | 100 | 36SMFG | 52 | ND | 80 | 8.2 | 23 | 3.6 (14) | 35 | 181 |
| RRLsin.cPPT.PGKGFP.Wpre (lenti) | VSV-G | 100 | S | 41 | ND | 36 | 0.4 (14) | 28 | ||||
| A00074 | RRLsin.cPPT.PGKYFP.Wpre (lenti) | VSV-G | 100 | 6SMF | 16 | ND | ND | 9.1 | 20 | 0.9 (9) | 26 | 177 |
| RRLsin.cPPT.PGKGFP.Wpre (lenti) | VSV-G | 10 | 6SMF | 6 | ND | ND | 0.1 (9) | 13 | ||||
| K99307 | RRLsin.cPPT.PGKGFP.Wpre (lenti) | VSV-G | 100 | 36SMFG | 17 | 26 | 55 | 19.7 | 20 | 1.1 (10) | 27 | 180 |
| MNDEYFPSN (onco) | Phoenix-GALV | 1 | 36SMFG | 50 | 26 | 70 | 23.2 (10) | 47 | ||||
| M99149 | RRLsin.cPPT.PGKGFP.Wpre (lenti) | VSV-G | 100 | 36SMFG | 6 | 33 | 40 | 17.6 | 20 | 0.3 (10) | 17 | 180 |
| MNDEYFPSN (onco) | Phoenix-GALV | 1 | 36SMFG | 34 | 29 | 55 | 12.1 (10) | 17 |
MOI indicates multiplicity of infection; S, SCF; M, MGDF; F, Flt3-L; G, G-CSF; ND, not done; ANC, absolute neutrophil count; lenti, lentiviral vector; and onco, oncoretroviral vector. In the column for growth factors, 3 indicates IL-3; and 6, IL-6.
The percentage of EGFP/EYFP-expressing leukocytes in the bulk liquid culture 3 to 5 days after transduction as determined by flow cytometry.
The total number of colonies per 1000 cells plated immediately after transduction.
The percentage of colonies PCR-positive for EGFP/EYFP, indicating the percentage of transduced CFU-Cs in the graft at the time of transplantation.