T cells cloned from CFSE cultures are antigen-specific
Original antigen/sorted cell type . | Cutoff CPM, mean + 2 SD* . | Con A†(cloning efficiency, %) . | Irrelevant antigen (cloning efficiency, %)† . | Same antigen (cloning efficiency, %)† . | Specificity, %‡ . |
|---|---|---|---|---|---|
| MBP mix/CD4+ cells | 17 947.50 | 3/12 (25) | 0/12 (0) | 9/24 (38) | 100 |
| MBP mix/CD8+ cells | 21 033.16 | 9/12 (75) | 0/12 (0) | 13/24 (54) | 72 |
| GA/CD4+ cells | 17 330.29 | 4/12 (33) | 0/12 (0) | 10/24 (42) | 100 |
| GA/CD8+ cells | 16 817.68 | 6/12 (50) | 0/12 (0) | 13/24 (54) | 100 |
| TT/CD4+ cells | 23 222.28 | 8/12 (67) | 0/12 (0) | 11/24 (46) | 69 |
| TT/CD8+ cells | 16 469.92 | 2/12 (17) | 0/12 (0) | 4/24 (17) | 100 |
Original antigen/sorted cell type . | Cutoff CPM, mean + 2 SD* . | Con A†(cloning efficiency, %) . | Irrelevant antigen (cloning efficiency, %)† . | Same antigen (cloning efficiency, %)† . | Specificity, %‡ . |
|---|---|---|---|---|---|
| MBP mix/CD4+ cells | 17 947.50 | 3/12 (25) | 0/12 (0) | 9/24 (38) | 100 |
| MBP mix/CD8+ cells | 21 033.16 | 9/12 (75) | 0/12 (0) | 13/24 (54) | 72 |
| GA/CD4+ cells | 17 330.29 | 4/12 (33) | 0/12 (0) | 10/24 (42) | 100 |
| GA/CD8+ cells | 16 817.68 | 6/12 (50) | 0/12 (0) | 13/24 (54) | 100 |
| TT/CD4+ cells | 23 222.28 | 8/12 (67) | 0/12 (0) | 11/24 (46) | 69 |
| TT/CD8+ cells | 16 469.92 | 2/12 (17) | 0/12 (0) | 4/24 (17) | 100 |
The cutoff to determine positive wells was calculated as mean counts per minute (CPM) of background (no antigen) wells (n = 12) plus 2 SD.
The numbers represent the proportion of wells that showed a proliferative response (CPM > cutoff) to the indicated stimulus. The response to Con A is representative of the cloning efficiency (survival of single-sorted T cells through rounds of PHA/IL-2 stimulation).
This number represents the calculated antigenic specificity of CFSE-sorted T cells (normalized to cloning efficiency; with a maximum value of 100%).