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Abstract
Ginsan, a Panax ginseng polysaccharide that contains glucopyranoside and fructofuranoside, has immunomodulatory effects. Although several biologic studies of ginsan have been performed, its effects on dendritic cells (DCs), which are antigen-presenting cells of the immune system, have not been studied. We investigated the immunomodulatory effects of ginsan on DCs. Ginsan had little effect on DC viability, even when used at high concentrations. Ginsan markedly increased the levels of production by DCs of IL-12 and TNF-α, as measured by ELISA. To examine the maturation-inducing activity of ginsan, we measured the surface expression levels of the maturation markers MHC class II and CD86 (B7.2) on DCs. It is interesting that ginsan profoundly enhanced the expression of CD86 on DC surfaces, whereas it increased that of MHC class II only marginally. In 3H-thymidine incorporation assays, ginsan-treated DCs stimulated significantly higher proliferation of allogeneic CD4+ T lymphocytes than did medium-treated DCs. Taken together, our data demonstrate that ginsan stimulates DCs by inducing maturation. Because DCs are critical antigen-presenting cells in immune responses, this study provides valuable information on the activities of ginsan.
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Fig. 1.
Viability of DCs treated with ginsan. DCs were seeded at 5×104 cells/well in 96-well culture plates and treated with different concentrations of ginsan for 2 days. After treatment, an MTT assay was performed, as described in Methods. The representative data, shown as mean±SD, are from three independent experiments that gave similar results. ∗∗p<0.01, ∗∗∗p<0.001.
Fig. 2.
Production of NO and cytokines by DCs treated with ginsan. For the NO assay (A), DCs were established in culture as described in Fig. 1. LPS or ginsan was added to the wells, and the culture supernatants were harvested 2 days later. The NO assay was performed as described in Methods. For cytokine measurements (B~D), DCs were seeded at 5×10 cells/well in 24-well culture plates and treated for 48 h. Concentrations of IL-12 (B), IL-10 (C), and TNF-α (D) in the culture media were measured using ELISA. In D, 1 μg/ml anti-mouse CD40 mAb, 1 μg/ml LPS, and 10 μg/ml ginsan were used. ∗p<0.05, ∗∗p<0.01, ∗∗∗p<0.001.
Fig. 3.
Ginsan increases the expression of maturation markers on the surfaces of DCs. DCs were established in culture and treated as described in Fig. 2B~D. Data shown are representative values from three separate experiments. ContDCs, medium alone- treated DCs; CD40DCs, DCs treated with 1 μg/ml anti-mouse CD40 mAb; GinsanDCs, DCs treated with 10 μg/ml ginsan.
Fig. 4.
Increased antigen-presenting capacities of ginsan-treated DCs. DCs were established in culture as described in Methods. For the MLR assay, DCs were co-cultured with 1×105 cells/well allogeneic CD4+ lymphocytes for 5 days. As described in Methods, the incorporated radioactivity was measured using a scintillation counter. The mean±SD level of radioactivity was derived from five individual wells (A). For the cytokine assay (B, C), 5×103 cells/well DCs were cocultured with 1×105 cells/well allogeneic CD4+ lymphocytes. The culture supernatants were harvested and used to measure IFN-γ (B) and IL-4 (C) levels. Shown are representative data from three independent experiments that gave similar results. ∗p<0.05, ∗∗p<0.01, ∗∗∗p< 0.001.
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