Fig. 3.

Dendritic cells isolated from castrated mice have differential effects on naïve (CD62L⁻ and antigen-experienced (CD62L⁺) CD4⁺ T cells. Irradiated DC isolated from sham-castrated (n=4) or castrated (n=6) C57BL/6 mice were cocultured with MACS-purified naïve or antigen-experienced CD4⁺ T cells from DBA/2 mice at a stimulator to responder ratio of 1:5 for 5 days. The cells were washed and re-plated to yield 5×10⁴ cells/well and non-specifically stimulated with PMA and calcium ionophore for 5 hours. The supernatant was collected and a Th₁/Th₂ multiplex cytokine assay carried out to determine if the DC skewed the Th₁/Th₂ phenotype of the naïve or antigen-experienced CD4⁺ T cells. A: Coculture of naïve CD62L⁺ CD4⁺ T cells DC isolated from castrated mice resulted in reduced cytokine expression. B: Coculture of antigen-experienced CD62L⁻ CD4⁺ T cells DC isolated from castrated mice resulted in increased cytokine expression. Figures are representative of two independent experiments. Data were analyzed using two-tailed t-tests. Abbreviations: DC; dendritic cell, MACS; magnetic-activated cell separation, PMA; phorbol 12-myristate 13-acetate.