The interferon lambda (IFN-l) cytokines have well-known antiviral properties, yet their contribution to immune regulation is not well understood. Epithelial cells represent the major target cell of IFN-l; peripheral blood mononuclear cells are generally considered nonresponsive, with the exception of plasmacytoid dendritic cells (pDCs). In this study we aimed to define the potential for discrete subpopulations of cells to directly respond to IFN-l. Analysis of peripheral blood leukocytes reveals that, while pDCs uniformly express the highest levels of IFN-l receptor, a small proportion of B cells and monocytes also express the receptor. Nevertheless, B cells and monocytes respond poorly to IFN-l stimulation in vitro, with minimal STAT phosphorylation and interferonstimulated gene (ISG) induction observed. We confirm that pDCs respond to IFN-l in vitro, upregulating their expression of pSTAT1, pSTAT3, and pSTAT5. However, we found that pDCs do not upregulate pSTAT6 in response to IFN-l treatment. Our results highlight unique aspects of the response to IFN-l and confirm that while the IFN-l receptor is expressed by a small proportion of several different circulating immune cell lineages, under normal conditions only pDCs respond to IFN-l stimulation with robust STAT phosphorylation and ISG induction. The difference in STAT6 responsiveness of pDCs to type I and type III interferons may help explain the divergence in their biological activities.