PV, PMF, and post-PV MF CD34+ cells contain JAK2V617F-positive SCID repopulating cells
| Patient no. . | CD34+ cell source . | JAK2V617F in granulocytes, % . | No of mice per sample . | No of CD34+ cells injected . | 6 weeks human cells, % . | 12 weeks human cells, % . | Analysis 15 weeks after transplantation . | ||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Human cells RF, % . | Human cells BM, % . | GpA+ cells, % . | CD33+ cells, % . | CD19+ cells, % . | JAK2V617F colonies over all colonies [% (no.)] . | JAK2V617F/total JAK2 in human CD45+, % . | |||||||
| PV1 | BM | 69 | 3 | 0.8 × 105 | 41.2 | 11.1 | 7.7 | 15.6 | 0 | 9.0 | 82.0 | NA (0% at 12 wk) | NA |
| PV2 | BM | 60 | 3 | 0.6 × 105 | 2.0 | 4.2 | 2.1 | 3.2 | 0 | 14.3 | 53.7 | NA (0% at 12 wk) | NA |
| PV3 | BM | 60 | 2 | 0.8 × 105 | 10.7 | 1.8 | 1.0 | 4.0 | 0 | 10 | 50.4 | NA | NA |
| PV4 | BM | 44 | 2 | 1.3 × 105 | 16.3 | 8.5 | 3.0 | 7.8 | 0 | 0.0 | 85.0 | 4 (2/47) | NA |
| PV5 | BM | 50 | 1 | 3.3 × 105 | 1.5 | 0.6 | 1.0 | 4.0 | 0 | 10.4 | 68.0 | 44 (4/9) | NA |
| PV6 | BM | 94 | 1 | 1.6 × 105 | 7.3 | 1.8 | 0.4 | 0.2 | 0 | NA | NA | 0 (0/1) | 3 |
| PV7 | BM | 57 | 1 | 1.6 × 105 | 18.4 | 34 | 18.1 | 24.7 | 0.1 | 20.0 | 77.0 | 0 (0/2) | 0 |
| PV8 | G-CSF mobilized PB | 40 | 1 | 6.2 × 105 | 8.0 | 20.5 | 59.0 | 58.0 | 0.6 | 53.0 | 42.0 | 9 (3/34) | 5 |
| PV9 | BM | 26 | 1 | 3.1 × 105 | 9.2 | 40.0 | 50.6 | 35.9 | 0.5 | 93.0 | 1.0 | 18 (14/79) | 1 |
| PMF1 | PB | 50 | 2 | 6.8 × 105 | 0.7 | 4.3 | 26.6 | 43.9 | 5 | 33.0 | 0 | 100 (181/181) | 67 |
| PMF2 | PB | 90 | 1 | 10 × 105 | 46.6 | 36.5 | 74.1 | 80.0 | 0.7 | 93.0 | 4.0 | 86 (60/70) | 76 |
| PMF3 | PB | NA | 1 | 1.4 × 105 | 1.3 | 0.5 | 1 | 0.8 | 0 | NA | NA | NA | 55 |
| PMF4 | PB | 50 | 1 | 5 × 105 | NA | 0.5 | 0.6 | 1.1 | 0 | 70.0 | 5.0 | 88 (7/8) | NA |
| PMF5 | PB | 30 | 2 | 5.6 × 105 | 2.2 | 0.2 | 0.7 | 0.7 | 0 | NA | NA | NA | 46 |
| PMF6 | PB | 50 | 1 | 6.5 × 105 | 0.1 | 0.2 | 0.8 | 1.0 | 0 | 40.0 | 0 | NA | 47 |
| PPV-MF1 | PB | 94 | 1 | 7.5 × 105 | 1.2 | 0 | 0.1 | 0.2 | 0 | NA | NA | NA | NA |
| PPV-MF2 | PB | NA | 1 | 6 × 105 | 0.4 | 1.2 | 0.6 | 0.4 | 0 | lNA | NA | NA | 95 |
| PPV-MF3 | PB | 100 | 1 | 4.7 × 105 | 3.1 | 1.4 | 1.4 | 1.4 | 0 | 80.0 | 2.0 | NA | 96 |
| PPV-MF4 | PB | 92 | 1 | 2.2 × 105 | 0 | 0.1 | 0 | 0.0 | 0 | NA | NA | NA | NA |
| Patient no. . | CD34+ cell source . | JAK2V617F in granulocytes, % . | No of mice per sample . | No of CD34+ cells injected . | 6 weeks human cells, % . | 12 weeks human cells, % . | Analysis 15 weeks after transplantation . | ||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Human cells RF, % . | Human cells BM, % . | GpA+ cells, % . | CD33+ cells, % . | CD19+ cells, % . | JAK2V617F colonies over all colonies [% (no.)] . | JAK2V617F/total JAK2 in human CD45+, % . | |||||||
| PV1 | BM | 69 | 3 | 0.8 × 105 | 41.2 | 11.1 | 7.7 | 15.6 | 0 | 9.0 | 82.0 | NA (0% at 12 wk) | NA |
| PV2 | BM | 60 | 3 | 0.6 × 105 | 2.0 | 4.2 | 2.1 | 3.2 | 0 | 14.3 | 53.7 | NA (0% at 12 wk) | NA |
| PV3 | BM | 60 | 2 | 0.8 × 105 | 10.7 | 1.8 | 1.0 | 4.0 | 0 | 10 | 50.4 | NA | NA |
| PV4 | BM | 44 | 2 | 1.3 × 105 | 16.3 | 8.5 | 3.0 | 7.8 | 0 | 0.0 | 85.0 | 4 (2/47) | NA |
| PV5 | BM | 50 | 1 | 3.3 × 105 | 1.5 | 0.6 | 1.0 | 4.0 | 0 | 10.4 | 68.0 | 44 (4/9) | NA |
| PV6 | BM | 94 | 1 | 1.6 × 105 | 7.3 | 1.8 | 0.4 | 0.2 | 0 | NA | NA | 0 (0/1) | 3 |
| PV7 | BM | 57 | 1 | 1.6 × 105 | 18.4 | 34 | 18.1 | 24.7 | 0.1 | 20.0 | 77.0 | 0 (0/2) | 0 |
| PV8 | G-CSF mobilized PB | 40 | 1 | 6.2 × 105 | 8.0 | 20.5 | 59.0 | 58.0 | 0.6 | 53.0 | 42.0 | 9 (3/34) | 5 |
| PV9 | BM | 26 | 1 | 3.1 × 105 | 9.2 | 40.0 | 50.6 | 35.9 | 0.5 | 93.0 | 1.0 | 18 (14/79) | 1 |
| PMF1 | PB | 50 | 2 | 6.8 × 105 | 0.7 | 4.3 | 26.6 | 43.9 | 5 | 33.0 | 0 | 100 (181/181) | 67 |
| PMF2 | PB | 90 | 1 | 10 × 105 | 46.6 | 36.5 | 74.1 | 80.0 | 0.7 | 93.0 | 4.0 | 86 (60/70) | 76 |
| PMF3 | PB | NA | 1 | 1.4 × 105 | 1.3 | 0.5 | 1 | 0.8 | 0 | NA | NA | NA | 55 |
| PMF4 | PB | 50 | 1 | 5 × 105 | NA | 0.5 | 0.6 | 1.1 | 0 | 70.0 | 5.0 | 88 (7/8) | NA |
| PMF5 | PB | 30 | 2 | 5.6 × 105 | 2.2 | 0.2 | 0.7 | 0.7 | 0 | NA | NA | NA | 46 |
| PMF6 | PB | 50 | 1 | 6.5 × 105 | 0.1 | 0.2 | 0.8 | 1.0 | 0 | 40.0 | 0 | NA | 47 |
| PPV-MF1 | PB | 94 | 1 | 7.5 × 105 | 1.2 | 0 | 0.1 | 0.2 | 0 | NA | NA | NA | NA |
| PPV-MF2 | PB | NA | 1 | 6 × 105 | 0.4 | 1.2 | 0.6 | 0.4 | 0 | lNA | NA | NA | 95 |
| PPV-MF3 | PB | 100 | 1 | 4.7 × 105 | 3.1 | 1.4 | 1.4 | 1.4 | 0 | 80.0 | 2.0 | NA | 96 |
| PPV-MF4 | PB | 92 | 1 | 2.2 × 105 | 0 | 0.1 | 0 | 0.0 | 0 | NA | NA | NA | NA |
A dose of 0.6 to 10 × 105 CD34+ cells per NOD/SCID mouse was transplanted. Six and 12 weeks after transplantation, the BM was aspirated from the right femur (RF) and analyzed for the degree of human cell engraftment (CD45+ cells). Fifteen weeks after transplantation, mice were killed and the previously aspirated RF was analyzed separately from the BM. Analysis of multilineage differentiation potential distinguished erythroid (Glycophorin A+), B-lymphoid (CD45+CD19+), and myeloid (CD45+CD33+) lineages. The mouse BM cells were seeded in methylcellulose, and human progenitors were picked and genotyped to look for the presence of the JAK2V617F mutation. DNA was also extracted from total BM cells, and the proportion of JAK2V617F over total human JAK2 was quantified.
NA indicates not available.