Ex vivo enumeration and phenotypic analysis of EBV-specific CTLs using major histocompatibility complex–peptide tetramers
| Donor code . | CD8 and tetramer* (% positive) . | CD27† (% positive/negative) . | CD38† (% positive/negative) . | CD44† (% positive/negative) . | CD62L† (% positive/negative) . | |||||
|---|---|---|---|---|---|---|---|---|---|---|
| GLC . | RPP . | GLC . | RPP . | GLC . | RPP . | GLC . | RPP . | GLC . | RPP . | |
| Patient 1 acute IM | 2.40 | 0.23 | 87/13 | 80/20 | 100/0 | 100/0 | 100/0 | 100/0 | 47/53 | 44/56 |
| After IM (11 mo) | 0.26 | 0.27 | 35/65 | 43/57 | 65/35 | 58/42 | 96/4 | 100/0 | 37/63 | 37/63 |
| Patient 2 acute IM | 4.40 | 0.61 | 63/37 | 63/37 | 76/24 | 87/13 | 96/4 | 93/7 | 21/79 | 35/65 |
| After IM (5 mo) | 0.72 | 0.41 | 33/67 | 32/68 | 68/32 | 70/30 | 94/6 | 100/0 | 50/50 | 47/53 |
| After IM (17 mo) | 1.14 | 0.23 | 41/59 | 28/72 | 63/37 | 37/63 | 96/4 | 86/14 | 17/83 | 10/90 |
| Donor 3 | 0.15 | 0.03 | 25/75 | 12/88 | 46/54 | 44/56 | 100/0 | 100/0 | 18/82 | 0/100 |
| Donor code . | CD8 and tetramer* (% positive) . | CD27† (% positive/negative) . | CD38† (% positive/negative) . | CD44† (% positive/negative) . | CD62L† (% positive/negative) . | |||||
|---|---|---|---|---|---|---|---|---|---|---|
| GLC . | RPP . | GLC . | RPP . | GLC . | RPP . | GLC . | RPP . | GLC . | RPP . | |
| Patient 1 acute IM | 2.40 | 0.23 | 87/13 | 80/20 | 100/0 | 100/0 | 100/0 | 100/0 | 47/53 | 44/56 |
| After IM (11 mo) | 0.26 | 0.27 | 35/65 | 43/57 | 65/35 | 58/42 | 96/4 | 100/0 | 37/63 | 37/63 |
| Patient 2 acute IM | 4.40 | 0.61 | 63/37 | 63/37 | 76/24 | 87/13 | 96/4 | 93/7 | 21/79 | 35/65 |
| After IM (5 mo) | 0.72 | 0.41 | 33/67 | 32/68 | 68/32 | 70/30 | 94/6 | 100/0 | 50/50 | 47/53 |
| After IM (17 mo) | 1.14 | 0.23 | 41/59 | 28/72 | 63/37 | 37/63 | 96/4 | 86/14 | 17/83 | 10/90 |
| Donor 3 | 0.15 | 0.03 | 25/75 | 12/88 | 46/54 | 44/56 | 100/0 | 100/0 | 18/82 | 0/100 |
PBMCs from IM patients 1 and 2 and healthy donor 3 were costained with anti–human CD8− tricolor-labeled antibody and phycoerythrin-labeled HLA A2/GLC or HLA B7/RPP tetramers. Fluorescence intensities were then measured with a FACScan. Values indicate the percent CD8 T cells positive for HLA A2/GLC or HLA B7/RPP tetramer.
For phenotypic analysis PBMCs from these individuals were costained with HLA-peptide tetramers for early lytic (GLC) or latent (RPP) epitopes and monoclonal antibodies specific to T-cell surface antigens (CD27, CD38, CD44, or CD62L). Values shown indicate the percent HLA-peptide tetramer–reactive T cells positive/negative for CD27, CD38, CD44, or CD62L.