New protein PCiP from edible golden oyster mushroom (Pleurotus citrinopileatus) activating murine macrophages and splenocytes

A new immunomodulatory protein (PCiP) was purified from an edible golden oyster mushroom (Pleurotus citrinopileatus) by extraction with 5% (v/v) cold acetic acid in the presence of 0.1% (v/v) 2‐mercaptoethanol, followed by ammonium sulfate fractionation, DE‐52 and MonoQ anion‐exchange chromatography. Electrophoresis assays demonstrated that the molecular mass of PCiP was approximately 15.0 kDa and its pI was around 5.2. PCiP is a simple protein without carbohydrate, and cannot agglutinate mouse red blood cells, suggesting PCiP is not a lectin. In addition, PCiP (5–20 µg mL−1) alone activated murine splenocytes, and markedly increased their proliferation and gamma‐interferon (IFN‐γ secretion, but suppressed MTT metabolization, while murine splenocytes were simultaneously stimulated by the mitogen concanavalin A (ConA). Furthermore, PCiP (5–20 µg mL−1) directly activated murine macrophages and increased the production of both the nitric oxide (NO) and tumor necrosis factor‐alpha (TNF‐α by RAW 264.7 macrophages. These findings suggest that PCiP could strengthen both the innate and adaptive responses of its host.