´╗┐Further studies will be required to analyze the ginsenoside composition of the RGEs and to verify which ginsenoside(s) contributes to the selective induction of IgA production

´╗┐Further studies will be required to analyze the ginsenoside composition of the RGEs and to verify which ginsenoside(s) contributes to the selective induction of IgA production. Open in a separate window Figure 1 Effects of red ginseng extract on B cell proliferation and antibody production. medicine for thousands of years to treat various diseases and to maintain body homeostasis. Many reports show that ginseng has multifunctional biological effects in immune function, anti-inflammatory, anti-cancer, anti-oxidant, metabolic processes (anti-diabetic) and the neuro-endocrine and cardiovascular system (blood pressure regulation) (1,2,3,4,5,6). Ginseng contains many active ingredients including ginsenosides, polysaccharides, peptides, phytosterols, polyacetylenic alcohols and fatty acids (2,4,7). Among them, ginsenosides are known to have the most pharmacological and immunological activity (4,8). In the case of Korean ginseng, 38 ginsenosides have been identified and classified into three groups: protopanaxadiol (PPD), protopanaxatriol (PPT) and oleanane (4). Recent investigations have exhibited that ginsenosides are responsible for regulation of the immune response. It has been reported that ginsenoside Rg1 regulates the innate immune response in dendritic cells and macrophages by differentially modulating the production of inflammatory cytokines (9,10). Rg1 also increases CD4+ T cell activity and modulates Th1/Th2 differentiation in vitro and in vivo (11,12). In addition, ginsenoside Rb1, Rd and Re elicit a Rabbit polyclonal to HspH1 Th1 and Th2 immune response (13,14,15,16), and recent studies have exhibited that these ginsenosides (Rg1, Rb1, Rd, Re and Rg3) have immunological adjuvant activity to enhance the immune response (17,18,19,20,21,22,23). Mature IgM+ B cells undergo Ig class switch recombination (CSR) at the switch region around the heavy chain locus to produce other Ig isotypes (IgG, IgA and IgE) and this class switching is usually selectively induced by cytokines such as IL-4, Peramivir IFN- and TGF-1 (24). In addition, expression of germline transcripts (GLTs) for each switch region is usually a prerequisite for each Ig CSR process (25). That is, the expression of GL transcripts induces IgA CSR. As mentioned above, ginsenosides act as adjuvants and then elicit both a humoral antibody response and a T cell mediated immune response. However, the direct effects of ginsenosides around the B cell response have not yet been investigated. To address this, we purified B cells from mouse splenocytes and examined the effects of reddish ginseng extract (RGE) and ginsenosides on B cell proliferation, antibody production, and expression of GLTs in vitro. Our study reveals that ginsenoside Rg1 and 20(S)-Rg3 selectively induce IgA production and GLT expression by LPS-activated mouse B cells. MATERIALS AND METHODS Animals BALB/c mice were purchased from Damool Science (Daejeon, Korea) and managed on an 8:16 h light:dark cycle in an animal environmental control chamber. Eight- to twelve-week-old mice were used, and animal care was in accordance with the institutional guidelines of the Institutional Animal Care and Use Committee of Konyang University or college. Purification of B cells, cell Peramivir culture, and reagents Mouse splenic B cells were purified by positive selection of B220+ cells using anti-B220 microbeads or by depletion of CD43+ cells using anti-CD43 microbeads and high-gradient magnetic cell separation (MACS; Miltenyi Biotec, Bergisch Gladbach, Germany) according to manufacturer’s instructions. Briefly, BALB/c mouse spleen cell suspensions were washed with HBSS (WelGENE, Daegu, Korea) and treated with 0.83% ammonium chloride to lyse the red blood cells. Spleen cells were treated with either anti-mouse B220 microbeads or anti-mouse CD43 microbeads and separated using a LS column and MACS Separator (Miltenyi Biotec, Peramivir Auburn, CA, USA). The purity of B cells (98%) was assessed by FACSCalibur (BD Biosciences, San Jose, CA, USA) after staining the cells with anti-CD43 FITC (eBioscience, San Diego, CA, USA) and/or anti-B220 PE (BD Biosciences). Cells were cultured at 37 in a humidified CO2 incubator (Forma Scientific, Marietta, OH, USA) in RPMI-1640 medium (WelGENE) supplemented with 10% fetal bovine serum (PAA Laboratories, Etobicoke, ON, Canada). Purified B cells were stimulated with Peramivir LPS (1 g/ml, InvivoGen, San Diego, CA, USA; 12.5 g/ml, Sigma-Aldrich, St Louis, MO, USA), red ginseng extract (200 g/ml, Prepared by Dr. JE Choi,.