Human Vα24⁺ Vβ11⁺ natural killer T cells (NKT cells) are “natural memory” T cells that detect glycolipid antigens such as α‐galactosylceramide (α‐GalCer) presented on CD1d. In the present study we found that highly purified Vα24⁺ NKT cells lack TLR9 mRNA, and thus are not sensitive towards stimulation with CpG oligodeoxynucleotides (ODN). Within PBMC, however, CpG ODN synergistically activated NKT cells stimulated with their cognate antigen α‐GalCer. Depletion of plasmacytoid dendritic cells (PDC) or myeloid dendritic cells (MDC) revealed that both DC subsets were necessary for the synergistic activation of NKT cells by α‐GalCer and CpG ODN. While PDC were responsible for the stimulation of NKT cells with CpG ODN, MDC but not PDC presented α‐GalCer via CD1d. Partial activation of NKT cells was mediated by PDC‐derived IFN‐α, whereas full activation of NKT cells as indicated by IFN−γ production required cell‐to‐cell contact of PDC and NKT cells in addition to IFN‐α; OX40 was involved in this interaction. We conclude that CpG‐activated PDC enhance α‐GalCer‐specific NKT cell activation, and bias activated NKT cells towards a Th1 phenotype. Our results lead to a novel concept of PDC function: to regulate effector activity of antigen‐stimulated T cells in a cell contact‐dependent manner without the need of simultaneous presentation of the cognate T cell antigen.