Immunostimulatory Effects of Complex GPB through Increased TNF‑α Expression and NK Cell Activation

Background: Complex Great Powerful Blend (GPB) (a great powerful blend) is a hot water extract of three well‑known dried fruits: goji berry, mulberry, and jujube. These fruits have been used as food for a long time, and this article sought to confirm the immunostimulatory effects of Complex GPB. Materials and Methods: To do this, in vitro study using RAW264.7 cells and in vivo using normal mice were performed. Cytokine production and NK cell activity levels were measured by enzyme‑linked immunosorbent assay, real‑time polymerase chain reaction (PCR), and the YAC‑1 cell‑killing assay. Transcription factor activation levels were also determined by western blotting analysis. Results: First, it was confirmed that Complex GPB had no cytotoxic effects on the cells and increased the production levels of tumor necrosis factor (TNF)‑a in a concentration‑dependent manner. Additionally, Complex GPB enhanced pro‑inflammatory cytokine expressions such as TNF‑a, IL‑12a, and IL‑1b. Western blotting analysis revealed that activation of the NF‑kB and AP‑1 pathways are involved. Furthermore, in vivo study confirmed that TNF‑a was found to be elevated in the serum and splenocytes of mice‑fed Complex GPB. Cytotoxic activation of NK cells was also increased in mice injected with Complex GPB. Conclusion: These results suggest that Complex GPB has an immune‑enhancing effect, which can be used for immunomodulatory remedies.