Figure 3.

Expansion of CD8⁺ CTLs by UV-irradiated RPMI 8226 MM cell whole-cell immunogens in 'mixed culture' conditions. Normal donor PBMCs were primed using UV-irradiated (120 mJ/cm²) or mitomycin-treated (100 μg/mL) RPMI 8226 MM cells as whole cell immunogens for 10 days and analyzed for CD4, CD8, CD56, granzyme and perforin expression using indirect immunofluorescence flow cytometric analysis. Unprimed and concurrently cultured PBMCs were used as negative controls. Dually positive granzyme⁺perforin⁺ CTLs were identified gated and specifically analyzed for CD4⁺ (open columns), CD8⁺ (black columns) or CD8⁺/CD56⁺ (striped columns) expression.