Extracts of cones and leaves of different species of the genus Juniperus have been used for centuries to treat a variety of medical problems; however, little is known about the active components conferring therapeutic properties to these extracts. To address this issue, we extracted water-soluble polysaccharides from Juniperus scopolorum cones and used ion exchange and size exclusion chromatography to separate them into five fractions, with estimated Mr of 30, 60, 100, 200, and 680 kDa, respectively. All fractions contained type II arabinogalactan in their structure, as determined by reaction with Yariv reagent and structural analysis by proton nuclear magnetic resonance spectroscopy, but lacked complement fixing activity. Analysis of the effects of Juniper polysaccharides on murine peritoneal macrophages, cultured J774.A1 macrophages, and human mononuclear phagocytes demonstrated that the high molecular weight polysaccharide fractions (200 and 680 kDa) had potent immunomodulatory activity. These polysaccharide fractions primed macrophages for an enhanced respiratory burst, directly stimulated NO production via induction of nitric oxide synthase, and induced macrophages to secrete both inflammatory (IL-1, IL-6, TNF-alpha, and IL-12) and anti-inflammatory (IL-10) cytokines. These data suggest that at least part of the beneficial therapeutic effects reported for extracts of juniper cones are due to modulation of monocyte/macrophage immune functions.