EBV and CMV peptide-specific cytotoxicity in HIV-infected and healthy donors requires perforin expression on tetramer-positive CD8 T cells: testing of immunomagnetically enriched tetramer-positive cells
| Subject . | HIV status . | Tetramer . | Tetramer-positive CD8 T cells, % . | Perforin+, % . | Specific cytotoxicity after selection . | |||
|---|---|---|---|---|---|---|---|---|
| Before selection . | After selection . | E/T ratio . | Tetramer E/T ratio . | % . | ||||
| 307 | Seropositive | A2 CMV | 0.96 | 79.6 | 25 | 2:1 | 1.6:1 | 15* |
| 501 | Seropositive | A2 CMV | 2.47 | 61.2 | 35 | 4.5:1 | 2.8:1 | 16* |
| CW16 | Seropositive | A2 EBV | 0.76 | 16.2 | 71 | 10:1 | 1.6:1 | 1.6 |
| HD2 | Seronegative | A2 CMV | 0.59 | 18.0 | 2.5 | 10:1 | 1.8:1 | 2.9 |
| HD3 | Seronegative | A2 EBV | 0.05 | 59.9 | 0 | 0.6:1 | 0.4:1 | 2.8 |
| Subject . | HIV status . | Tetramer . | Tetramer-positive CD8 T cells, % . | Perforin+, % . | Specific cytotoxicity after selection . | |||
|---|---|---|---|---|---|---|---|---|
| Before selection . | After selection . | E/T ratio . | Tetramer E/T ratio . | % . | ||||
| 307 | Seropositive | A2 CMV | 0.96 | 79.6 | 25 | 2:1 | 1.6:1 | 15* |
| 501 | Seropositive | A2 CMV | 2.47 | 61.2 | 35 | 4.5:1 | 2.8:1 | 16* |
| CW16 | Seropositive | A2 EBV | 0.76 | 16.2 | 71 | 10:1 | 1.6:1 | 1.6 |
| HD2 | Seronegative | A2 CMV | 0.59 | 18.0 | 2.5 | 10:1 | 1.8:1 | 2.9 |
| HD3 | Seronegative | A2 EBV | 0.05 | 59.9 | 0 | 0.6:1 | 0.4:1 | 2.8 |
PBMCs were tested after immunomagnetic enrichment for tetramer-positive cells for cytotoxicity against autologous B-cell lines incubated with tetrameric peptide or media. The frequencies of tetramer-positive cells before and after selection are shown, together with the percentages of tetramer-positive cells that stain for perforin. Tetramer E/T ratio gives the ratio of peptide-specific cells to peptide-loaded targets, using the frequency of tetramer-positive cells after enrichment. *Percent specific cytotoxicity is greater than 5%.