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?4D and E). These data suggest that ginsenosides induce cytokine secretion via the activation of phosphorylated ERK1/2 (pERK1/2) and phosphorylated JNK (pJNK) signaling BMS-345541 in CD14+ monocytes. Fig.?4 Activation of ERK1/2 and JNK signaling in lipopolysaccharide (LPS)-sensitized CD14+ monocytes with or without ginsenoside fractions. (A) The CD14+ monocytes were stimulated with LPS (50?ng/mL) for 30?min or with ginsenoside fractions (10?��g/mL) ... 3.4. Ginsenosides attenuate the differentiation of CD14+ monocytes into DCs Monocytes differentiate into DCs when cultured in the presence of GM-CSF and IL-4 [8]. To test whether ginsenoside fraction is involved in DC differentiation, CD14+ monocytes were incubated with GM-CSF and IL-4 in the presence or absence of ginsenoside fractions for 3 d or 5 d, and the expression of cell surface and maturation markers (i.e., CD80, CD86, CD40, CD11c, CD14, and MHC class II) was measured [9]. Three days after the treatment, little to no change had occurred (Fig.?5A). However, 5 d after the treatment, the ginsenoside fractions suppressed the expression of CD80, CD86, CD40, and CD11c, but not MHC class II and CD14 (Fig.?5B). These results indicate that DCs treated with ginsenoside fractions during the maturation process express low levels of costimulatory molecules. Fig.?5 Suppression of the expression of cell surface molecules during dendritic cell differentiation in the presence or absence of ginsenoside fractions. The CD14+ monocytes were treated Megestrol Acetate with interleukin 4 (IL-4; 500 U/mL) and granulocyte macrophage colony-stimulating ... 3.5. DCs differentiated in the presence of ginsenoside fractions exhibit impaired maturation status and T cell activation Mature DCs express higher levels of surface markers such as CD80, CD86, CD40, and CD83, compared to immature DCs [14]. Therefore, to further examine the characteristics of DCs differentiated in the presence of ginsenoside fractions (Gin-DCs), the Gin-DCs were treated with LPS. To identify the impact of Gin-DCs on the maturation process, we measured the expression of the surface markers CD80, CD86, CD40, Histone Methyltransferase inhibitor and MHC class II. As Fig.?6A shows, the expression of these markers decreased in a dose-dependent manner, whereas the expression of CD40 remained relatively unchanged. Fig.?6 Dendritic cell (DC) maturation in the presence of ginsenoside fractions and the regulation of CD4+ T cells. (A) The DCs treated with ginsenoside fractions (the unbroken, dotted, and broken lines indicate 0 ��g/mL, 1 ��g/mL, and 10 ��g/mL, ... To investigate whether Gin-DCs activate CD4+ T cells, the Gin-DCs were primed for 2 d with ethanol-killed S.?aureus [12]. They were then cocultured with CFSE-labeled CD4+ T cells for an additional 3 d or 5 d. The CD4+ T cell proliferation was slightly suppressed when cocultured with S.?aureus-primed Gin-DCs for 5 days (Fig.?6B). In addition, IFN-�� production decreased significantly (p?
