Yield of P495 tetramer+ CTLs and cytotoxicity after a single stimulation with ER− PBMCS, BLCLs, and AAPCs
APC . | Total number of T cells after coculture (× 10−6) . | Percentage of HLA A2.1/P495 tetramer-positive cells . | Absolute number of P495 tetramer-positive cells*(× 10−6) . | Percent killing of T2 cells†(E/T ratio 10:1) . | Absolute number of P495 tetramer+ cells obtained from 100 mL of blood (× 10−6) . |
|---|---|---|---|---|---|
| Donor A | |||||
| ER− PBMC | 1.8 ± 0.08 | 22.7 ± 2.5 | 0.42 ± 0.06 | 52 ± 5.3 | 6.67 ± 1.01 |
| BLCL | 1.9 ± 0.16 | 18.0 ± 4.6 | 0.34 ± 0.09 | 44 ± 14.3 | 16.19 ± 4.10 |
| AAPCA2P495 | 2.1 ± 0.18 | 11.0 ± 2.6 | 0.23 ± 0.08 | 61 ± 5.5 | 10.96 ± 3.61 |
| AAPCA2pp65 | 1.8 ± 0.25 | 35.7 ± 3.8 | 0.63 ± 0.03 | 68 ± 3.5 | 30.23 ± 1.24 |
| Donor J | |||||
| ER− PBMC | 2.0 ± 0.08 | 15.7 ± 1.2 | 0.32 ± 0.02 | 31 ± 4.0 | 7.58 ± 0.57 |
| BLCL | 2.2 ± 0.13 | 6.3 ± 0.6 | 0.09 ± 0.01 | 19 ± 5.0 | 6.49 ± 0.45 |
| AAPCA2P495 | 2.2 ± 0.28 | 14.7 ± 1.5 | 0.32 ± 0.06 | 41 ± 8.0 | 22.47 ± 4.05 |
| AAPCA2pp65 | 1.5 ± 0.13 | 31.3 ± 3.1 | 0.45 ± 0.01 | 55 ± 5.0 | 31.62 ± 0.44 |
| Donor Y | |||||
| ER− PBMC | 1.6 ± 0.10 | 14.3 ± 1.2 | 0.23 ± 0.03 | 38 ± 4.8 | 7.73 ± 0.90 |
| BLCL | 2.7 ± 0.13 | 11.0 ± 2.0 | 0.30 ± 0.05 | 29 ± 2.3 | 20.85 ± 3.38 |
| AAPCA2P495 | 1.2 ± 0.19 | 13.3 ± 1.5 | 0.17 ± 0.04 | 30 ± 4.0 | 11.36 ± 3.12 |
| AAPCA2pp65 | 1.6 ± 0.06 | 28.3 ± 4.2 | 0.44 ± 0.05 | 46 ± 4.0 | 30.96 ± 3.38 |
APC . | Total number of T cells after coculture (× 10−6) . | Percentage of HLA A2.1/P495 tetramer-positive cells . | Absolute number of P495 tetramer-positive cells*(× 10−6) . | Percent killing of T2 cells†(E/T ratio 10:1) . | Absolute number of P495 tetramer+ cells obtained from 100 mL of blood (× 10−6) . |
|---|---|---|---|---|---|
| Donor A | |||||
| ER− PBMC | 1.8 ± 0.08 | 22.7 ± 2.5 | 0.42 ± 0.06 | 52 ± 5.3 | 6.67 ± 1.01 |
| BLCL | 1.9 ± 0.16 | 18.0 ± 4.6 | 0.34 ± 0.09 | 44 ± 14.3 | 16.19 ± 4.10 |
| AAPCA2P495 | 2.1 ± 0.18 | 11.0 ± 2.6 | 0.23 ± 0.08 | 61 ± 5.5 | 10.96 ± 3.61 |
| AAPCA2pp65 | 1.8 ± 0.25 | 35.7 ± 3.8 | 0.63 ± 0.03 | 68 ± 3.5 | 30.23 ± 1.24 |
| Donor J | |||||
| ER− PBMC | 2.0 ± 0.08 | 15.7 ± 1.2 | 0.32 ± 0.02 | 31 ± 4.0 | 7.58 ± 0.57 |
| BLCL | 2.2 ± 0.13 | 6.3 ± 0.6 | 0.09 ± 0.01 | 19 ± 5.0 | 6.49 ± 0.45 |
| AAPCA2P495 | 2.2 ± 0.28 | 14.7 ± 1.5 | 0.32 ± 0.06 | 41 ± 8.0 | 22.47 ± 4.05 |
| AAPCA2pp65 | 1.5 ± 0.13 | 31.3 ± 3.1 | 0.45 ± 0.01 | 55 ± 5.0 | 31.62 ± 0.44 |
| Donor Y | |||||
| ER− PBMC | 1.6 ± 0.10 | 14.3 ± 1.2 | 0.23 ± 0.03 | 38 ± 4.8 | 7.73 ± 0.90 |
| BLCL | 2.7 ± 0.13 | 11.0 ± 2.0 | 0.30 ± 0.05 | 29 ± 2.3 | 20.85 ± 3.38 |
| AAPCA2P495 | 1.2 ± 0.19 | 13.3 ± 1.5 | 0.17 ± 0.04 | 30 ± 4.0 | 11.36 ± 3.12 |
| AAPCA2pp65 | 1.6 ± 0.06 | 28.3 ± 4.2 | 0.44 ± 0.05 | 46 ± 4.0 | 30.96 ± 3.38 |
The total number of T cells per 1 × 106 responder cells, the percentage of P495 tetramer-positive cells, and the absolute number of P495 tetramer-positive cells from 100 mL of peripheral blood are shown for 3 donors (A, J, Y). Values represent mean ± standard deviation after 10 to 14-day coculture on ER− PBMCs, BLCL, AAPCA2P495, and AAPCA2pp65. Mean values represent at least 3 experiments.
Number of P495 tetramer-positive cells = [total number of T cells] × [% of tetramer-positive cells]:
Specific killing of T2 cells = [% killing of T2 cells pulsed with P495] minus [% killing of T2 cells pulsed with flu].