Figure 4.
Treatment with temsirolimus has minimal effect on baseline expression of DC markers; however, temsirolimus decreases the capacity of DCs to stimulate T-cell proliferation. (A) To assess baseline DC markers, BM-derived DCs were treated in vitro for 24 h with varying doses of temsirolimus. The DCs were examined by flow cytometry while gating on CD11c. (B) To assess DC function, BM-derived DCs were stimulated with vaccine (complex of hsp110 and gp100) and then treated with temsirolimus (50 ng ml−1). These DCs were then used to stimulate CD8 T cells purified by negative selection from Pmel-1 lymphocytes. Proliferation of Pmel-1 T cells was assessed by monitoring CSFE dilution while gating on CD8. Representative results are shown from at least three independent experiments.