Immune Netw.  2016 Apr;16(2):140-145. 10.4110/in.2016.16.2.140.

In vitro Stimulation of NK Cells and Lymphocytes Using an Extract Prepared from Mycelial Culture of Ophiocordyceps sinensis

• Jang SH
• Park J
• Jang SH
• Chae SW
• Jung SJ
• So BO
• Ha KC
• Sin HS
• Jang YS

• KMID: 2168055
• DOI: http://doi.org/10.4110/in.2016.16.2.140

Abstract

Ophiocordyceps sinensis is a natural fungus that has been valued as a health food and used in traditional Chinese medicine for centuries. The fungus is parasitic and colonizes insect larva. Naturally occurring O. sinensis thrives at high altitude in cold and grassy alpine meadows on the Himalayan mountain ranges. Wild Ophiocordyceps is becoming increasingly rare in its natural habitat, and its price limits its use in clinical practice. Therefore, the development of a standardized alternative is a great focus of research to allow the use of Ophiocordyceps as a medicine. To develop an alternative for wild Ophiocordyceps, a refined standardized extract, CBG-CS-2, was produced by artificial fermentation and extraction of the mycelial strain Paecilomyces hepiali CBG-CS-1, which originated from wild O. sinensis. In this study, we analyzed the in vitro immune-modulating effect of CBG-CS-2 on natural killer cells and B and T lymphocytes. CBG-CS-2 stimulated splenocyte proliferation and enhanced Th1-type cytokine expression in the mouse splenocytes. Importantly, in vitro CBG-CS-2 treatment enhanced the killing activity of the NK-92MI natural killer cell line. These results indicate that the mycelial culture extract prepared from Ophiocordyceps exhibits immune-modulating activity, as was observed in vivo and this suggests its possible use in the treatment of diseases caused by abnormal immune function.

Keyword

O. sinensis; Immune modulation; Mycelial extract; NK cell

MeSH Terms

Altitude
Animals
Colon
Ecosystem
Fermentation
Food, Organic
Fungi
Homicide
Insects
Killer Cells, Natural*
Larva
Lymphocytes*
Medicine, Chinese Traditional
Mice
Paecilomyces
T-Lymphocytes

Figure

• 

  Figure 1 Level of splenocyte proliferation induced by various concentrations of in vitro CBG-CS-2 treatment together with (A) T cell mitogen, Con A and (B) B cell mitogen, LPS. Stimulation indices were calculated by dividing the CPM of the sample from the CBG-CSG-2 treatment with that of the control non-treated sample. The CPM of the control non-treated sample was (A) 75,298.67±1997.07 and (B) 8,857.67±242.61.

• 

  Figure 2 Stimulation of Th1-type cytokine expression by in vitro stimulation of splenocytes with CBG-CS-2. Levels of IFN-γ in the splenocyte culture supernatant after in vitro stimulation with the indicated concentration of CBG-CS-2 or placebo were determined in the presence of 1 µg/ml ConA. ***p<0.001 indicates a significant difference in the results obtained.

• 

  Figure 3 Stimulating activity of in vitro treatment of CBG-CS-2 on NK-92MI-mediated cytotoxicity against K562 target cells. (A) NK-92MI cells were stimulated with CBG-CS-2 for 144 hrs and incubated with 51 Cr-labelled K562 target cells with an E:T ratio of 200:1. A similar pattern of cytotoxicity was observed when E:T ratios of 100:1 and 50:1 were used (data not shown). (B) NK-92MI cells were stimulated with either CBG-CS-2 or a placebo for 144 hrs and incubated with 51 Cr-labelled K562 target cells with indicated E:T ratios. ***p<0.001 indicates a significant difference in the results obtained.

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