Korean J Physiol Pharmacol.  2011 Apr;15(2):101-106. 10.4196/kjpp.2011.15.2.101.

Characterization and Immunopotentiating Effects of the Glycoprotein Isolated from Dioscorea Batatas

• Huong PT
• Lee CH
• Li MH
• Lee MY
• Kim JK
• Lee SM
• Seon JH
• Lee DC
• Jeon YJ

• KMID: 2285414
• DOI: http://doi.org/10.4196/kjpp.2011.15.2.101

Abstract

We demonstrate that glycoprotein isolated from Dioscorea batatas (GDB) has immunostimulatory effects including macrophage activation. Analysis of infiltration of inflammatory cells into peritoneal cavity showed GDB treatment significantly increased the recruitment of macrophages, lymphocytes, neutrophils, and monocytes into the peritoneal cavity. Treatment of spleen cells isolated from C57BL/6 mice with GDB significantly increased the proliferation of B cells and T cells induced by LPS and ConA, respectively. Treatment with GDB significantly increased the cytolytic capacity of NK cells and macrophages against YAC-1 and B16 cells, respectively. In order to further confirm and investigate the mechanism of GDB on macrophage activation, we analyzed the effects of GDB on the cytokine expression including iNOS, IL-1beta, and TNF-alpha in mouse macrophage cell line, RAW 264.7 cells. RT-PCR and ELISA showed that GDB increased the expression of IL-1beta, and TNF-alpha, whereas iNOS was not induced by GDB. Collectively, this series of experiments indicates that GDB stimulates immune system including macrophage activation.

Keyword

Glycoprotein; Macrophages; TNF-alpha; IL-1beta

MeSH Terms

Animals
B-Lymphocytes
Cell Line
Dioscorea
Enzyme-Linked Immunosorbent Assay
Glycoproteins
Immune System
Killer Cells, Natural
Lymphocytes
Macrophage Activation
Macrophages
Mice
Monocytes
Neutrophils
Peritoneal Cavity
Spleen
T-Lymphocytes
Tumor Necrosis Factor-alpha
Glycoproteins
Tumor Necrosis Factor-alpha

Figure

• 

  Fig. 1. Characterization of glycoprotein isolated from Dioscorea batatas. (A) The lyophilized proteins were dissolved in 50 mM Tris-HCl buffer (pH 8.0) and subjected to an anion-exchange chromatography. Fractionation of proteins was performed by a step-wise salt gradient (0.1, 0.2, 0.3, and 1 M) NaCl in equilibrium buffer. (B) Peak fractions were further analyzed by SDS-PAGE.

• 

  Fig. 2. Effect of GDB on (A) LPS- and (B) ConA-stimulated splenic lymphocyte proliferation. Splenocytes were isolated from C57BL/6 mice. The results are presented as the mean±S.E.M. ∗∗Denote significant differences (p<0.05, p<0.01) vs. the control group. Experiments were repeated three times.

• 

  Fig. 3. Effects of GDB on (A) NK cell- and (B) macrophage-mediated cytotoxicity. Cytotoxicity was measured as described in Methods and was expressed as the cytolytic percentage of target cells. IL-2 and LPS/IFN-γ were used as a positive control of NK cell- and macrophage-mediated cytotoxicity, respectively. The results are presented as the mean±S.E.M. ∗∗Denotes significant differences (p <0.01) vs. the control group. Experiments were repeated three times.

• 

  Fig. 4. Effects of GDB on macrophage activation. (A) RAW264.7 cells were treated with GDB for 8 h. Total RNA was then isolated and analyzed for iNOS, IL-0, and TNF-α using RT-PCR. The production of (B) NO2–, (C) TNF-α, and (D) IL-1β were analyzed from the cell lysates treated with GDB for 24 h. The results are presented as the mean± S.E.M. ∗∗Denotes significant differences (p<0.01) vs. the control group. One representative of three experiments is shown.

Reference

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