A novel water-soluble polysaccharide was identified in the pupae of the melon fly (Bactrocera cucurbitae) as a molecule that activates the mammalian innate immune response. We attempted to purify this innate immune activator using nitric oxide (NO) production in mouse RAW264 macrophages as an indicator of immunostimulatory activity. A novel acidic polysaccharide was identified, which we named “dipterose”, with a molecular weight of 1.01×106 and comprising nine monosaccharides. Dipterose was synthesized in the melon fly itself at the pupal stage. The NO-producing activity of dipterose was approximately equal to that of lipopolysaccharide, a potent immunostimulator. Inhibition of Toll-like receptor 4 (TLR4) led to the suppression of NO production by dipterose. Furthermore, dipterose induced the expression of proinflammatory cytokines and interferon β (IFNβ) and promoted the activation of nuclear factor kappa B (NF-κB) in macrophages, indicating that it stimulates the induction of various cytokines in RAW264 cells via the TLR4 signaling pathway. Our results thus suggest that dipterose activates the innate immune response against various pathogenic microorganisms and viral infections. This is the first identification of an innate immune-activating polysaccharide from an animal.

We have identified a novel acidic polysaccharide from silkmoth (Antheraea yamamai) pupae that activates the mammalian innate immune response. This bioactive polysaccharide was isolated using nitric oxide production in mouse RAW264 macrophages as an indicator of immunostimulatory activity. We named this polysaccharide "silkrose". It has a molecular weight of 3.15 × 10(5) and comprises nine monosaccharides. The expression profiles indicated that silkrose induced the expression of proinflammatory cytokines and interferon β that exist downstream of MyD88-dependent and MyD88-indeptendent signaling pathways. Also, the inhibition of Toll-like receptor 4 (TLR4), which exists upstream of the signaling pathways, led to the suppression of NO production by silkrose. Furthermore, this polysaccharide promoted the activation of nuclear factor kappa B in RAW264 cells, indicating that it stimulates the induction of various cytokines in macrophages through the TLR4 signaling pathway. Our results thus suggest that silkrose activates the innate immune response to various pathogenic microorganisms and viral infections.

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