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. Author manuscript; available in PMC: 2008 Jun 30.
Published in final edited form as: J Biol Chem. 2005 Apr 18;280(30):27491–27501. doi: 10.1074/jbc.M500555200

Fig. 3. The effects of differing pMHCI/CD8 interactions on tetramer staining of 868 CTL.

Fig. 3

The CTL line 868 is derived from an HIV-1 infected donor and is specific for the HLA A2-restricted p17 Gag epitope SLYNTVATL. 2 × 105 868 cells per test were resuspended in 20 μl of PBS and stained with D227K/T228A HLA A2 (277/8KA), A245V HLA A2 (A245V), wild type HLA A2 (wildtype), Q115E HLA A2 (Q115E), or A2/Kb α3 domain fusion (A2/Kb) tetramers folded around the SLYNTVATL peptide at either 0.5, 5 or 50 μg/ml for 20 min at 37 °C. Cells were then stained with CD8-allophycocyanin (clone SK1) for 30 min on ice, washed twice, and then resuspended in PBS. Data were acquired using a FACSCalibur flow cytometer and analyzed using CellQuest software. All HLA A2-SLYNTVATL tetramer-positive cells in the 868 CTL line bear a Vα12-2, Vβ5-6 TCR of identical sequence.