Anti cd8 apc sk1

Example 3

The following reagents were used: anti-CD3 FITC (Clone BW264/56, Miltenyi Biotec), anti-CD8-APC (SK1, BD Pharmingen), Dextramer-HPV E7_11-20-PE (Dext-HPV PE)(HLA-A*0201; YMLDLQPETT, Immudex), Dextramer-HIV-1 P17 Gag 77-85-PE (Dext-HIV-PE)(HLA-A*0201; SLYNTVATL, Immudex), anti-TCR-Vß3-FITC (JOVI-3, Ancell). Cells were incubated with Dext as recommended by the manufacturer for 10 min at room temperature, then mAb were added for 30 min at 4° C. Stained cells were washed with PBS/0.5% BSA/0.1% NaN3 (PBA), and were analyzed using a FACSCalibur flow cytometer (Becton Dickinson).

Example 15

Anti-CD3 FITC (Clone BW264/56, Miltenyi Biotec), anti-CD8-APC (SK1, BD Pharmingen), Dextramer-HPV E7_11-20-PE (Dext-HPV PE; HLA-A*0201; YMLDLQPETT, Immudex), Dextramer-HIV-1 P17 Gag 77-85-PE (Dext-HIV-PE; HLA-A*0201; SLYNTVATL, Immudex), anti-TCR-Vß3-FITC (JOVI-3, Ancell) were used. Cells were incubated with dextramer for 10 min at room temperature, then anti-CD3, -CD8, -Vß3 antibodies were added for 30 min at 4° C. Perforin and granzyme intracellular staining was performed with specific antibodies in PBS/0.5% BSA/0.1% NaN3 containing saponin (0.5%) at room temperature for 20 min. Stained cells were washed with PBS/0.5% BSA/0.1% NaN3 and analyzed using a FACSCalibur flow cytometer (Becton Dickinson).