Figure 4.

Dose-response curve of Treg induction in CTCL cells. A. Increasing numbers of CD3-treated apoptotic CTCL cells (0.3 × 10⁶, 1.5 × 10⁶, 3.0 × 10⁶) were added to a constant number of DC (10⁵), co-cultured overnight and used to stimulate freshly isolated autologous CTCL cells. As controls, increasing numbers of non-loaded DC were cultured with CD4-treated viable CTCL cells. Then additional CTCL cells were added and co-cultured overnight. The percentage of membrane CD3⁺/cytoplasmic CTLA-4⁺ CTCL cells was measured by flow cytometry gated on the lymphocyte population. B. Increasing numbers of apoptotic CTCL cells (0.10 × 10⁶, 0.5 × 10⁶, 1.0 × 10⁶) were added to 10⁵ DC and co-cultured overnight, followed by addition of 10⁶ purified CTCL cells. The percentage of membrane CD4⁺/cytoplasmic FoxP3⁺ expressing cells was measured by flow cytometry. Apoptotic = CD3-treated, Viable = CD4-treated