Table 3.
Composition of T cells and DCs obtained via apheresis before and after Treg enrichment and after expansion culture under GMP
| Mononuclear cells | DC* (n=3) | Treg | |||
|---|---|---|---|---|---|
| Before column (n = 3) | After column (n = 3) | Before culture† (n = 3) | After culture (n = 4) | ||
| Viable | |||||
| N × 106 | 94 (57.2-127.8) | 27 718 (21 778-37 000) | 257.2 (139-477) | 101.7 (65-146.9) | 69.9 (50.5-97.2) |
| % | 88 (79-99.1) | 99.9 (99.9-100) | 99.9 (99.8-100) | 94.5 (90.7-97.7) | 88.8 (72.5-99.3) |
| CD4+CD127−CD25+ Foxp3+ | |||||
| N × 106 | ND | 799.6 (424.6-1265) | 206.6 (114.5-386.1) | 87.7 (55.2-132.9) | 63.36 (41.5-97.1) |
| % | ND | 2.8 (1.9-3.4) | 79.9 (76.4-82.4) | 80.5 (76.3-83) | 78.3 (72.5-85.2) |
| CD4+CD127−CD25+ Foxp3− | |||||
| N × 106 | ND | 234.5 (34-581.8) | 10.9 (1.5-22.2) | 2.7 (0.3-5.7) | 2.8 (0.2-6.2) |
| % | ND | 0.7 (0.1-1.6) | 3.8 (1.1-5.8) | 3.1 (0.2-5.8) | 3.5 (0.2-6.8) |
| CD4+CD127−CD25− | |||||
| N × 106 | ND | 1082 (369.4-1821) | 3.6 (1.2-5.7) | 2.0 (0.1-3.4) | 1.0 (0.7-1.4) |
| % | ND | 3.6 (1.7-4.9) | 1.5 (0.9-2.5) | 1.6 (0.1-2.5) | 1.3 (1.1-1.4) |
| CD4+CD127+ | |||||
| N × 106 | 11.4 (0.5-22.8)‡ | 9871 (3248-20 370) | 17.6 (9.0-31.3) | 6.2 (4.6-7.8) | 5.8 (3.3-8.3) |
| % | 10.3 (0.7-23.3) | 32.0 (13.3-55.1) | 7.0 (6.5-8) | 6.2 (3.8-8.0) | 6.9 (6.0-9.1) |
| CD8+ | |||||
| N × 106 | 3.2 (0.5-6.4) | 5205 (684-2562) | 5.2 (1.0-13.3) | 1.7 (0.3-3.5) | 1.2 (0.4-2.9) |
| % | 2.5 (0.8-4) | 17.1 (9.3-26.3) | 1.4 (0.7-2.8) | 1.3 (0.6-2.2) | 1.2 (0.8-2.2) |
| CD19+ | |||||
| N × 106 | 14 (0.1-24.6) | 1704 (684.5-2562) | 7.1 (6.1-9.0) | 2.4 (0.5-4.4) | 1.7 (0.8-2.7) |
| % | 18.2 (0.1-37) | 6.2 (3.1-10.5) | 3.8 (1.3-5.8) | 2.2 (0.7-4.5) | 2.4 (0.6-3.2) |
| CD16+ or CD56+ | |||||
| N × 106 | 8.5 (6.6-12.2) | 2362 (8.7-3,589) | 0.4 (0-1.0) | 0.1 (0-0.3) | 1.5 (0.2-4.3) |
| % | 9.8 (4.1-18.3) | 8 (0-14.3) | 0.1 (0-0.2) | 0.1 (0-0.3) | 1.4 (0.5-3.2) |
| CD14+ | |||||
| N × 106 | 16.9 (4.1-24.7) | 5836 (2087-8882) | 1.8 (0.1-4.8) | 0.6 (0.1-1.4) | 0.2 (0-0.5) |
| % | 20.2 (2.5-32.9) | 24 (5.6-36.4) | 0.4 (0.1-1) | 0.6 (0.1-1.4) | 0.2 (0-0.6) |
| CD14−CD11c+ | |||||
| N × 106 | 68 (29-132) | 2081 (1613-2812) | 0.7 (0.2-1.4) | 0.2 (0.1-0.4) | 1.3 (0.8-2.5) |
| % | 58.8 (30-81.6) | 7.5 (7.4-7.6) | 0.2 (0.1-0.3) | 0.3 (0.1-0.6) | 0.2 (1-1.9) |
Data are presented as mean (range).
*
DCs were γ-irradiated (3000 cGy) before coculture with Tregs to prevent contamination.
†
A fraction of the purified Tregs were cultured with DCs from an HLA-matched sibling at a 1:1 ratio with IL-2, IL-15, and rapamycin.
‡
This population was not tested for CD127 expression.