Figure 3. DC transfer induces CD8 and CD4 function in vivo.

Bone marrow derived DC were generated and cultured overnight in media with or without CpG ODN 1826 (10 μM) prior to peptide-pulsing and transfer via tail vein infusion at 2×10⁶ cells/mouse. (A) 2×10⁶ CFSE-labeled CD8-sorted P14/Thy1.1⁺ splenocytes were transferred to C57BL/6 mice alone or with a separate infusion of peptide-pulsed CpG-stimulated DCs. 3 days later, splenocytes were isolated and flow cytometry was conducted to determine CFSE dilution of transferred Thy1.1⁺CD8⁺ cells. (B) 2×10⁶ CFSE-labeled CD4-sorted Smarta/Thy1.1⁺ splenocytes were transferred to C57BL/6 mice alone or with a separate infusion of peptide-pulsed CpG-stimulated DCs. 3 days later, splenocytes were isolated and flow cytometry was conducted to determine CFSE dilution of transferred Thy1.1⁺CD4⁺ cells. (C) Quantification of gp33–41-specific production of IFN-γ, TNF-α, and IL-2 by CD8⁺ splenocytes in naïve mice and 8 days after vaccination with unstimulated or CpG-stimulated peptide-pulsed DCs. Results show means+/−S.D. Dot plots are gated on Thy1.1⁺ population. All results are representative of 2–3 independent experiments with >3 mice per group.