Evaluation of lentivectors for in vivo transduction of HSCs in humanized mice
| Mice . | Reconstitution, % hCD45+cells* . | BM, % GFP+ cells/total Lin+ cells† . | Thymus, % GFP+ cells† . | Spleen, % GFP+ cells† . | PB, % GFP+ cells† . | Vector titer, IU/mL . | |||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| BM . | Thymus . | Spleen . | PB . | h45+ . | h34+ . | h13+ . | h19+ . | h45+ . | h45+ . | h45+ . | |||
| RDTR/SCFHA 1 | 7 | 80 | 6 | F1 | 2.5 | 6.6 | 1 | 0.4 | 0 | 0 | 0 | 4E8 | |
| F2 | 2.3 | 6.5 | 6.6 | 0.3 | |||||||||
| RDTR/SCFHA 2 | 40 | 85 | 56 | 2 | F1 | 2.5 | 3.3 | 5 | 0.7 | 0 | 0.05 | 0 | 9E7 |
| F2 | 3.8 | 4 | 7.3 | 0.5 | |||||||||
| RDTR/SCFHA 3 | 18 | 98 | 47 | 7 | F1 | 2.2 | 4.3 | 6.6 | 0.06 | 0 | 0.006 | 0 | 1E8 |
| F2 | |||||||||||||
| RDTR/SCFHA 4 | 46 | 93 | 30 | 3 | F1 | 2.1 | 2.6 | 6.1 | 0.8 | 0 | 0.04 | 0 | 2.9E7 |
| F2 | 1.7 | 3 | 4.2 | 0.7 | |||||||||
| RDTR/SCFHA 5 | 40 | 99 | 15 | 5 | F1 | 1.9 | 2.2 | 6.8 | 0.7 | 0 | 0.06 | 0 | 4E7 |
| F2 | 2.8 | 4.2 | 4.5 | 0.7 | |||||||||
| RDTR/SCFHA 6 | 19 | 94 | 36 | 30 | F1 | 2 | 2.6 | 2.5 | 0.03 | 0 | 0.03 | 0 | 5E7 |
| F2 | 2.8 | 2.1 | 6.1 | 0.04 | |||||||||
| RDTR/SCFHA 7 | 6 | 97 | 5 | 1 | F1 | 1.2 | 4 | 2.9 | 0.8 | 0 | 0 | 0 | 2.9E7 |
| F2 | |||||||||||||
| Means | 2.3 | 3.8 | 4.9 | 0.4 | 0 | 0.02 | 0 | ||||||
| SD (±) | 0.6 | 1.5 | 1.9 | 0.3 | 0 | 0.02 | 0 | ||||||
| RDTR 1 | 10 | 91 | 30 | 4 | F1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1.3E8 |
| F2 | 0 | 0 | 0 | 0 | |||||||||
| RDTR 2 | 7 | 90 | 5 | 6 | F1 | 0.05 | 0 | 0 | 0.05 | 0 | 0 | 0 | 2E8 |
| F2 | 0.4 | 0 | 0 | 0.05 | |||||||||
| RDTR 3 | 15 | 93 | 20 | 11 | F1 | 0.6 | 0 | 0 | 0.6 | 0 | 0 | 0 | 2E8 |
| F2 | 0.8 | 0 | 0 | 0.8 | |||||||||
| RDTR 4 | 7 | 97 | 12 | 4 | F1 | 0.1 | 0 | 0 | 0.3 | 0 | 0 | 0 | 4E7 |
| F2 | 0.2 | 0 | 0 | 0.4 | |||||||||
| Means | 0.3 | 0 | 0 | 0.3 | 0 | 0 | 0 | ||||||
| SD (±) | 0.3 | 0 | 0 | 0.3 | 0 | 0 | 0 | ||||||
| RDTR 1+ SCFHA 1 | 21 | 92 | 18 | 10 | F1 | 0.04 | 0 | 0.01 | 0.03 | 0 | 0 | 0 | 1.3E8 |
| F2 | 0.01 | 0 | 0.00 | 0.01 | |||||||||
| RDTR 2+ SCFHA 2 | 17 | 95 | 11 | 4 | F1 | 0.02 | 0 | 0 | 0.03 | 0 | 0 | 0 | 2E8 |
| F2 | 0.2 | 0 | 0 | 0.25 | |||||||||
| RDTR 3+ SCFHA 3 | 5 | 86 | 7 | F1 | 0.0 | 0 | 0 | 0 | 0 | 0 | 0 | 4E7 | |
| F2 | 0.2 | 0 | 0 | 0.3 | |||||||||
| Mice . | Reconstitution, % hCD45+cells* . | BM, % GFP+ cells/total Lin+ cells† . | Thymus, % GFP+ cells† . | Spleen, % GFP+ cells† . | PB, % GFP+ cells† . | Vector titer, IU/mL . | |||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| BM . | Thymus . | Spleen . | PB . | h45+ . | h34+ . | h13+ . | h19+ . | h45+ . | h45+ . | h45+ . | |||
| RDTR/SCFHA 1 | 7 | 80 | 6 | F1 | 2.5 | 6.6 | 1 | 0.4 | 0 | 0 | 0 | 4E8 | |
| F2 | 2.3 | 6.5 | 6.6 | 0.3 | |||||||||
| RDTR/SCFHA 2 | 40 | 85 | 56 | 2 | F1 | 2.5 | 3.3 | 5 | 0.7 | 0 | 0.05 | 0 | 9E7 |
| F2 | 3.8 | 4 | 7.3 | 0.5 | |||||||||
| RDTR/SCFHA 3 | 18 | 98 | 47 | 7 | F1 | 2.2 | 4.3 | 6.6 | 0.06 | 0 | 0.006 | 0 | 1E8 |
| F2 | |||||||||||||
| RDTR/SCFHA 4 | 46 | 93 | 30 | 3 | F1 | 2.1 | 2.6 | 6.1 | 0.8 | 0 | 0.04 | 0 | 2.9E7 |
| F2 | 1.7 | 3 | 4.2 | 0.7 | |||||||||
| RDTR/SCFHA 5 | 40 | 99 | 15 | 5 | F1 | 1.9 | 2.2 | 6.8 | 0.7 | 0 | 0.06 | 0 | 4E7 |
| F2 | 2.8 | 4.2 | 4.5 | 0.7 | |||||||||
| RDTR/SCFHA 6 | 19 | 94 | 36 | 30 | F1 | 2 | 2.6 | 2.5 | 0.03 | 0 | 0.03 | 0 | 5E7 |
| F2 | 2.8 | 2.1 | 6.1 | 0.04 | |||||||||
| RDTR/SCFHA 7 | 6 | 97 | 5 | 1 | F1 | 1.2 | 4 | 2.9 | 0.8 | 0 | 0 | 0 | 2.9E7 |
| F2 | |||||||||||||
| Means | 2.3 | 3.8 | 4.9 | 0.4 | 0 | 0.02 | 0 | ||||||
| SD (±) | 0.6 | 1.5 | 1.9 | 0.3 | 0 | 0.02 | 0 | ||||||
| RDTR 1 | 10 | 91 | 30 | 4 | F1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1.3E8 |
| F2 | 0 | 0 | 0 | 0 | |||||||||
| RDTR 2 | 7 | 90 | 5 | 6 | F1 | 0.05 | 0 | 0 | 0.05 | 0 | 0 | 0 | 2E8 |
| F2 | 0.4 | 0 | 0 | 0.05 | |||||||||
| RDTR 3 | 15 | 93 | 20 | 11 | F1 | 0.6 | 0 | 0 | 0.6 | 0 | 0 | 0 | 2E8 |
| F2 | 0.8 | 0 | 0 | 0.8 | |||||||||
| RDTR 4 | 7 | 97 | 12 | 4 | F1 | 0.1 | 0 | 0 | 0.3 | 0 | 0 | 0 | 4E7 |
| F2 | 0.2 | 0 | 0 | 0.4 | |||||||||
| Means | 0.3 | 0 | 0 | 0.3 | 0 | 0 | 0 | ||||||
| SD (±) | 0.3 | 0 | 0 | 0.3 | 0 | 0 | 0 | ||||||
| RDTR 1+ SCFHA 1 | 21 | 92 | 18 | 10 | F1 | 0.04 | 0 | 0.01 | 0.03 | 0 | 0 | 0 | 1.3E8 |
| F2 | 0.01 | 0 | 0.00 | 0.01 | |||||||||
| RDTR 2+ SCFHA 2 | 17 | 95 | 11 | 4 | F1 | 0.02 | 0 | 0 | 0.03 | 0 | 0 | 0 | 2E8 |
| F2 | 0.2 | 0 | 0 | 0.25 | |||||||||
| RDTR 3+ SCFHA 3 | 5 | 86 | 7 | F1 | 0.0 | 0 | 0 | 0 | 0 | 0 | 0 | 4E7 | |
| F2 | 0.2 | 0 | 0 | 0.3 | |||||||||
Two- to 4-day-old newborn BALB/c Rag2−/−γc−/− mice were subjected to a sublethal irradiation of 2 × 1.5 Gy. 2 × 105 hCB CD34+ cells were injected intrahepatically into the newborns. After 13 weeks of reconstitution, the humanized mice were injected into the right (F1) and the left (F2) femur with 10 μL of the indicated vector stock. The animals were killed 2 weeks after intrafemoral injection of the vectors, and the BM, spleens, thymi, and PB were harvested and assessed for levels of human cell engraftment. Multilineage engraftment was demonstrated by Lin+ markers as indicated, and the percentage of EGFP+ cells was determined by FACS. Independent experiments were performed with different CD34+ CB samples and different preparations of each vector. Vector titers (IU/mL) were determined on 293T cells
Percentage of human cells (hCD45+ cells) per total number of human and mouse mononucleated cells in each hematopoietic tissue.
Percentage of GFP+ cells per total Lin+ cells (ie, % GFP+CD13+ cells/total CD13+ cells).