Survival of cultured polyclonal T cells stimulated with anti-CD3 and anti-CD28 after cytokine withdrawal.

Flow cytometric analysis of cultured T cells stained with FITC-coupled annexin V and PI. LV'VFas⁺ T cells cultured with anti-CD3 (B) or combined anti-CD3 and anti-CD28 (C) for 12 to 13 days were washed and plated on autologous γ-irradiated peripheral blood mononuclear cells as feeder cells in the absence of IL-2 or mitogen stimulation. Fresh PBLs (A) were plated in a similar fashion, and after 7 days cell viability was assessed by staining with annexin V and PI. T cells stimulated with anti-CD3 alone demonstrated a marked increase in apoptosis as compared with freshly isolated PBLs.