Table 1.
Phenotypic characterization and CMV-specific CTL isolation of unmanipulated PBMCs and the non-adherent fraction
| Unmanipulated PBMCs | Non-adherent fraction | p | |
|---|---|---|---|
| Leukocyte subpopulations | |||
| CD14+ cells | 47.8% (36.0 – 55.0) | 2.1% (1.3 – 6.1) | 0.005 |
| T lymphocytes (CD3+) | 24.0% (18.0 – 37.1) | 69.9% (31.9 – 79.2) | 0.007 |
| CTLs (CD3 + CD8+) | 17.7% (6.8 – 25.1) | 35.2% (23.8 – 45.2) | 0.009 |
| Helper T cells (CD3 + CD4+) | 12.5% (11.4 – 19.4) | 19.3% (16.6 – 29.6) | 0.208 |
| Viability (7-AAD-) | 98.0% (95.6 – 98.5) | 96.0% (93.4 – 97.8) | 0.066 |
| PM staining: specific and non-specific binding | |||
| CMV-specific CTLs (CD3 + CD8 + PM+) | 0.14% (0.06 – 0.62) | 0.65% (0.24 – 1.51) | 0.003 |
| Non-specific PM binding (CD8-PM+) | 0.56% (0.41 – 0.86) | 0.16% (0.12 – 0.37) | 0.003 |
| CMV-specific CTL isolation | |||
| Purity | 20.8% (6.9 – 61.7) | 76.0% (32.7 – 83.7) | 0.043 |
| Yield | 38.6% (32.9 – 44.8) | 42.1% (23.4 – 84.1) | 0.893 |
Cell subsets, CMV-specific CTLs and non-specific PM staining were analyzed in the unmanipulated PBMC sample and in the non-adherent cells (n = 11). CMV-specific CTLs were isolated from unmanipulated PBMCs and non-adherent fraction, and purity and yield of the obtained cellular product were determined (n = 5). Comparison was done with the Wilcoxon signed-rank test and significance level was fixed to p < 0.05.