Fig. 5

Effects of DNA vaccination with pHSP90 on dendritic cells (DCs). a Popliteal lymph node DCs from pHSP90-vaccinated mice (sorted as indicated in Supplementary Fig. 3A) were analyzed for cell surface expression of MHC class II and costimulatory molecules (CD40, CD80, and CD86). b Fold changes in median fluorescence intensity (MFI) of the same markers as a in DCs where HSP90 was either expressed as intracellular (i.c.) (following transfection with Chariot reagent) or extracellular (e.c.) protein (present in the culture medium of non-transfected DCs). *P < 0.05 in the comparisons with control untreated DCs (0–6 h). c Effects of intracellular vs. extracellular HSP90 expression in DCs on Tregs. Reduced expression of MHC class II and costimulatory molecules (CD40, CD80, CD86) in DCs expressing HSP90 intracellularly associated with the expansion of Tregs from CD4⁺CD25⁻ T cells (negatively sorted with magnetic beads) in cultures for 3 days at a 1:4 ratio with DCs. *P < 0.04 between control or e.c. HSP90 and i.c. HSP90. Dots in b and c represent individual points; columns show cumulative results + SEM