Table 2.
Online quality assessment of human CD4+ and CD8+ T cells
| Functiona,b,c | Markers | Day − 4d | Day 0 | Day 2 | Day 4 | Day 8 (re-stimulated) | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| CD4+ | CD8+ | CD4+ | CD8+ | CD4+ | CD8+ | CD4+ | CD8+ | CD4+ | CD8+ | ||
| Cell type | CD4+ | 99.1 ± 0.5 | 0.7 ± 0.0 | 99.0 ± 1.1 | 0.8 ± 0.0 | 99.6 ± 0.2 | 0.6 ± 0.3 | 99.6 ± 0.4 | 0.4 ± 0.2 | 98.6 ± 1.1 | 0.3 ± 0.0 |
| CD8+ | 0.8 ± 0.1 | 99.5 ± 0.0 | 0.4 ± 0.3 | 99.8 ± 0.0 | 0.3 ± 0.1 | 99.3 ± 0.0 | 0.4 ± 0.4 | 93.8 ± 0.0 | 0.4 ± 0.1 | 99.0 ± 0.1 | |
| Activation signal | CD3+ (stimulation) | 97.8 ± 2.6 | 98.2 ± 2.2 | 97.5 ± 2.8 | 92.6 ± 8.4 | 99.4 ± 0.7 | 96.3 ± 4.1 | 98.3 ± 2.3 | 91.7 ± 7.7 | 95.1 ± 3.7 | 96.8 ± 0.5 |
| ICOS+/CD278+ (co-stimulation) | 30.3 ± 19.9 | 13.3 ± 10.8 | 99.6 ± 0.1 | 99.5 ± 0.6 | 99.7 ± 0.2 | 99.2 ± 1.0 | 99.5 ± 0.6 | 91.2 ± 12.7 | 99.8 ± 0.2 | 99.8 ± 0.2 | |
| OX40+/CD134+ (co-stimulatione) | 1.8 ± 1.3 | 0.1 ± 0.1 | 2.7 ± 1.6 | 1.7 ± 1.3 | 0.2 ± 0.2 | 0.1 ± 0.1 | 0.3 ± 0.5 | 0.1 ± 0.1 | 0.4 ± 0.3 | 0.0 ± 0.0 | |
| CD27+ (co-stimulation) | 83.3 ± 2.7 | 64.1 ± 5.1 | 83.1 ± 9.9 | 95.4 ± 1.1 | 87.9 ± 4.7 | 86.6 ± 4.0 | 92.3 ± 3.4 | 89.7 ± 1.3 | 67.2 ± 2.4 | 88.5 ± 1.7 | |
| Inhibitory signal | PD-1+/CD279+ (TCR signaling) | 5.7 ± 4.3 | 6.5 ± 5.6 | 84.5 ± 4.3 | 84.7 ± 4.3 | 30.3 ± 24.6 | 7.0 ± 3.8 | 8.1 ± 3.9 | 3.3 ± 2.5 | 63.1 ± 6.0 | 67.9 ± 15.6 |
| LAG-3+/CD223+ | 95.0 ± 6.2 | 97.6 ± 2.8 | 23.3 ± 11.9 | 61.6 ± 15.8 | 11.0 ± 5.8 | 24.0 ± 12.7 | 4.4 ± 1.2 | 4.9 ± 4.0 | 16.4 ± 8.6 | 47.4 ± 8.6 | |
| KLRG1+ | 0.9 ± 0.4 | 1.4 ± 0.7 | 5.5 ± 2.5 | 7.9 ± 2.3 | 7.5 ± 0.7 | 6.6 ± 1.0 | 6.7 ± 2.3 | 6.4 ± 3.4 | 6.4 ± 1.5 | 4.7 ± 0.4 | |
| Memory typef | CCR7+ (central memory T cell) | 19.4 ± 7.6 | 6.0 ± 1.0 | 90.7 ± 6.6 | 86.5 ± 8.0 | 78.3 ± 5.8 | 70.1 ± 5.2 | 81.7 ± 5.5 | 68.5 ± 6.4 | 39.1 ± 9.9 | 29.2 ± 7.2 |
| CD45RO+ (memory T cell) | 86.7 ± 4.0 | 57.4 ± 13.6 | 97.0 ± 4.4 | 99.7 ± 0.4 | 98.4 ± 1.9 | 93.5 ± 7.8 | 98.6 ± 0.9 | 94.1 ± 2.6 | 99.6 ± 0.3 | 99.0 ± 0.1 | |
| CD45RA+ (naïve T cell) | 13.9 ± 3.0 | 55.3 ± 6.6 | 2.4 ± 1.1 | 3.5 ± 2.6 | 3.7 ± 2.3 | 3.6 ± 2.4 | 0.7 ± 0.3 | 1.1 ± 0.7 | 1.9 ± 1.5 | 3.4 ± 0.9 | |
Notes
aThe CD4+ T cells and CD8+ T cells were cultivated in single culture. The T cells used for data collection came from 3 donors.
bFlow cytometry was used to access T cell quality and the expression of these markers was reported as the binding intensity of antibodies.
cAll data are presented as mean ± standard error of the mean (SEM).
dDay-4 data was collected from isolated cells without stimulation. Day 8 data was collected after 2nd stimulation.
eCo-stimulation could happen with various mechanisms25. The low values of OX40/CD134 indicated the low possibility of this receptor correlated co-stimulation mechanism.
fAll cells obtained a memory phenotype after stimulation.