Although Cordyceps militaris is documented several medicinal functions, there is not enough for demonstration of leukocytic differentiation. Cordycepin and adenosine were 11.75 µg and 1.25 µg in the extract, respectively. Unlike the levels of TNF-α and IL-1β in macrophages that were approximately 4 time and 48 times higher than the control under lipopolysaccharides (LPS), macrophages under the extract (1 μg/mL) showed 13- and 10-fold lower TNF-α and IL-1β levels than the LPS-treated cells. This was corroborated by flow cytometry, where their levels were 20 times and 14 times lower, respectively. Under the extract, the LPS-treated macrophages enhanced M2 polarization and attenuated M1 polarization. In addition, the extract also dose-dependently activated macrophage phagocytosis. Under the extract conditioned medium, dendritic cells (DCs) were strongly differentiated toward CD11b+ and Xcr1+ cells because their densities were 13.6 times and 6.26 times higher than those in the LPS conditioned medium, respectively. Differentiation of Treg and natural killer T-like (NKTL) cells also were increased about 1.67 times and 6.73 times than those in the LPS conditioned medium, respectively. These results suggest that the C. militaris extract has strong effects on the modulation of macrophages and dendritic cells and T cells under inflammatory stress.

Backgrounds: Cordyceps militaris is a well-known medicinal fungus. Cordycepin, a metabolite of this fungus, has strong biological activities against leukemia, oxidative stress, aging, tumors, and inflammation. Methods: HPLC analysis was conducted to measure the content of corydycepin in the extract. Real time PCR was performed to evaluate the cytokines. Immunoactivity including the polarization, phagocytic activity and cellular differentiations were evaluated by flow cytometry. Results: The yields of cordycepin and adenosine in the extract were 11.75 µg and 1.25 µg (per gram fresh mycelium), respectively. From measurements of the bioactivity in the extract, the levels of TNF-α and IL-1β in macrophages treated with lipopolysaccharides (LPS) were found to be approximately 4 and 48 times higher than those in the control, as shown by qRT-PCR. However, cells treated with 1 µg/mL of the extract showed 13 and 10-fold lower TNF-α and IL-1β levels when compared to LPS-treated cells. This was corroborated by flow-cytometry, where their levels were 20 and 14 times lower, respectively. Addition of the extract to LPS-treated cells enhanced M2 polarization and attenuated M1 polarization. In addition, the extract also dose-dependently activated macrophage phagocytosis. Under treatment with the extract conditioned medium, DCs were strongly differentiated toward CD11b+ and Xcr1+ cells as their density were 13.6 and 6.26 times higher than those in the LPS conditioned medium, respectively. Moreover, the number of Treg and NKT cells differentiated in the extract conditioned medium were increased about 1.67 and 6.73 times than those in the LPS conditioned medium, respectively. Conclusions: These results suggest that the C. militaris hydrolytic extract has strong effects on the modulation of immune actors, such as macrophages and dendritic cells, under inflammatory stress.