Increases in the number of EBV-specific CD8⁺ cells over the course of polyclonal CTL line generation.

EBV-specific polyclonal CTL lines from HLA-A2 donors ES (A), SB (B), MS (C), and TZ (D) were established. Cells were harvested on day 28 and cryopreserved. On the day of the ELISPOT assay, aliquots of CTL lines were thawed and incubated with stimulators in multiscreen HA plate. Stimulators for ELISPOT assay included autologous LCLs (1 × 10⁵/well), and autologous PBMCs (1 × 10⁵/well) in medium containing the LMP2 or BMLF1 peptide (10 μg/mL, final concentration). The fold-increase in the number of LCL-responsive (filled diamonds), LMP2 peptide-responsive (open circles), and BMLF1 peptide-responsive CD8⁺ cells (open triangles) is indicated to the right of the day 28 data point.