Short-Term and Long-Term Reconstitution of Hematopoietic Lineages by Mobilized PB
| . | Short-Term Reconstitution (1 mo) . | |||||
|---|---|---|---|---|---|---|
| . | Source of Reconstituting Cells (no. cells injected) . | Control PB (5 × 105) . | FL PB . | FL PB . | FL + G-CSF PB . | FL + G-CSF PB . |
| . | Control BM (2 × 105) . | . | (5 × 105) . | (1 × 105) . | (5 × 105) . | (1 × 105) . |
| Lineage . | % Donor Phenotype (range) . | |||||
| . | . | . | . | . | . | . |
| T cell | 99 (97-100) | 0 (0) | 64 (44-88) | 48 (15-91) | 73 (32-97) | 47 (6-94) |
| B cell | 80 (74-88) | 1 (0-2) | 35 (22-57) | 24 (8-53) | 46 (44-49) | 39 (5-63) |
| Myeloid | 36 (19-63) | 0 (0-1) | 18 (16-21) | 26 (0-70) | 38 (6-62) | 12 (1-31) |
| . | Short-Term Reconstitution (1 mo) . | |||||
|---|---|---|---|---|---|---|
| . | Source of Reconstituting Cells (no. cells injected) . | Control PB (5 × 105) . | FL PB . | FL PB . | FL + G-CSF PB . | FL + G-CSF PB . |
| . | Control BM (2 × 105) . | . | (5 × 105) . | (1 × 105) . | (5 × 105) . | (1 × 105) . |
| Lineage . | % Donor Phenotype (range) . | |||||
| . | . | . | . | . | . | . |
| T cell | 99 (97-100) | 0 (0) | 64 (44-88) | 48 (15-91) | 73 (32-97) | 47 (6-94) |
| B cell | 80 (74-88) | 1 (0-2) | 35 (22-57) | 24 (8-53) | 46 (44-49) | 39 (5-63) |
| Myeloid | 36 (19-63) | 0 (0-1) | 18 (16-21) | 26 (0-70) | 38 (6-62) | 12 (1-31) |
| . | Long-Term Reconstitution (6 mo) . | |||||
|---|---|---|---|---|---|---|
| . | Control BM (2 × 105) . | Control PB (5 × 105) . | FL PB (5 × 105) . | FL PB (1 × 105) . | FL + G-CSF PB (5 × 105) . | FL + G-CSF PB (1 × 105) . |
| T cell | 76 (65-86) | 3 (3) | 48 (12-74) | 10 (5-14) | 14 (9-17) | 11 (6-14) |
| B cell | 88 (76-99) | 0 (0) | 55 (29-79) | 4 (3-5) | 36 (25-46) | 15 (7-19) |
| Myeloid | 74 (63-85) | 0 (0) | 63 (48-88) | 10 (7-14) | 39 (31-50) | 20 (12-28) |
| . | Long-Term Reconstitution (6 mo) . | |||||
|---|---|---|---|---|---|---|
| . | Control BM (2 × 105) . | Control PB (5 × 105) . | FL PB (5 × 105) . | FL PB (1 × 105) . | FL + G-CSF PB (5 × 105) . | FL + G-CSF PB (1 × 105) . |
| T cell | 76 (65-86) | 3 (3) | 48 (12-74) | 10 (5-14) | 14 (9-17) | 11 (6-14) |
| B cell | 88 (76-99) | 0 (0) | 55 (29-79) | 4 (3-5) | 36 (25-46) | 15 (7-19) |
| Myeloid | 74 (63-85) | 0 (0) | 63 (48-88) | 10 (7-14) | 39 (31-50) | 20 (12-28) |
C57BL/6 mice (Ly5.1+) were irradiated with 1,000 R, then injected IV 3 to 4 hours later with congenic donor cells (Ly5.2+). Control BM cells and PBMNC were isolated from donor mice treated for 10 days with MSA (1 μg/mouse/d). PBMNC were also collected from donor mice treated with MSA plus FL (10 μg/mouse/d) for 10 days or MSA plus FL and G-CSF (both at 10 μg/mouse/d) for 6 days. Ten to 15 recipient mice were injected with donor cells from each respective group. Three mice per group were harvested 30 days (short-term) or 6 months (long-term) postirradiation to evaluate donor reconstitution. For analysis of short-term reconstitution, only thymus (for T-cell staining) and BM (for B and myeloid cell staining) were harvested, whereas for analysis of long-term reconstitution, thymus, BM, and spleens were harvested and stained (splenic data shown above). T, B, and myeloid cells were detected with the MoAbs: Thy1.2, B220, and MAC1, respectively. Donor reconstitution was evaluated using flow cytometry where the above tissues were stained with lineage specific MoAbs and the A20 MoAb that recognizes donor cells (Ly5.2+). Limit of detection of donor cells was 1%. Short-term and long-term reconstitution potential from PBMNC from FL-treated mice was performed in two separate experiments each giving similar results.
N = 2.