Immune Netw.  2014 Oct;14(5):260-264. 10.4110/in.2014.14.5.260.

Immunomodulatory Effects of ZYM-201 on LPS-stimulated B Cells

• Lee YE
• Kim S
• Jung WJ
• Lee HS
• Kim MY

• KMID: 2391723
• DOI: http://doi.org/10.4110/in.2014.14.5.260

Abstract

ZYM-201 is a methyl ester of triterpenoid glycoside from Sanguisorba officinalis which has been used for treatment of inflammatory and metabolic diseases. In this study, immunomodulatory effects of ZYM-201 on B cells were examined in vitro and in vivo. When splenocytes were activated with lipopolysaccharide (LPS), the major population which had shown an increase in cell numbers was B cells. However, when the B cells were treated with ZYM-201 after LPS activation, their cell numbers and the expression of major costimulatory molecules, CD80 and CD86, were decreased. Furthermore, the effect of LPS, which induces activation of NF-kappaB, was abolished by ZYM-201: LPS-stimulated B cells showed decrease of phosphorylation after treatment of ZYM-201. The same results were shown in vivo experiments. These results suggest that ZYM-201 may play a role in the modulation of inflammatory responses through inhibiting NF-kappaB activation and downregulating the expression of costimulatory molecules on B cells.

Keyword

ZYM-201; B cell; Inflammation; LPS

MeSH Terms

B-Lymphocytes*
Cell Count
Inflammation
Metabolic Diseases
NF-kappa B
Phosphorylation
Sanguisorba
NF-kappa B

Figure

• 

  Figure 1 Effect of ZYM-201 on the activation of LPS-stimulated B cells in vitro. (A) and (B) Relative percentages of lymphocytes and B cells. Splenocytes were assessed after treatment with/without LPS (1 µg/ml) for 24 hours. In the case of ZYM-201 treatment, 50 and 100 µM ZYM-201 was added 6 hours after LPS treatment. Lymphocyte numbers were calculated by using SPHERO™ AccuCount Blank Particles (Spherotech, Lake Forest, IL) according to manufacturer's recommendations. Percentage of lymphocyte numbers was determined by comparison to control (100%). Percentage of B cells was decided using B220 mAbs and compared to control (100%). Error bars show standard deviation of six independent experiments. The statistical evaluation of the data was performed with Student's t-test and one-way analysis of variance. **p<0.01. (C) Flow cytometric analysis of CD80 and CD86 expression on B cells cultured with LPS and/or ZYM-201 (50 µM). Filled histograms show isotype-matched control Abs and numbers above bracketed line shows the percentage of positive cells for each Ab staining. Histograms are representative and mean fluorescence intensity (MFI) shown below each histogram is the average of six independent experiments. (D) Western blot analysis of levels of NF-κB p65 phosphorylation in B cells. Enriched B cells were cultured with LPS (1 µg/ml) and ZYM-201 (0 to 200 µM) for 30 minutes. Data shows total and phosporylated NF-κB p65 at serine536. HSP90 was used as loading control. Results are representative of six independent experiments.

• 

  Figure 2 in vivo analysis of ZYM-201 effects on B cells of LPS-injected mice. Mice were intraperitoneally injected with LPS (1 mg/kg) and/or orally administrated with ZYM-201 (20 mg/kg). Twenty four hours after LPS injection, spleens were taken. (A) and (B) Relative percentages of lymphocytes and B cells. The numbers of lymphocytes and B220+ B cells were calculated by using SPHERO™ AccuCount Blank Particles and the percentage of the cells was determined by comparison to control (100%). Error bars show standard deviation of four independent experiments. **p<0.01; ***p<0.005. (C) CD80 and CD86 expression on B cells. Filled histograms show isotype-matched control Abs and numbers above bracketed line shows the percentage of positive cells for each Ab staining. Histograms are representative and mean fluorescence intensity (MFI) shown below each histogram is the average of four independent experiments.

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