J Vet Sci.  2011 Jun;12(2):187-189. 10.4142/jvs.2011.12.2.187.

Maturation of bone marrow-derived dendritic cells by a novel beta-glucan purified from Paenibacillus polymyxa JB115

• Ko EJ
• Byon YY
• Jee Y
• Shin T
• Park SC
• Hahn TW
• Joo HG

• KMID: 1106562
• DOI: http://doi.org/10.4142/jvs.2011.12.2.187

Abstract

We investigated the immunostimulatory effects of a novel beta-glucan purified from Paenibacillus (P.) polymyxa JB115 on bone marrow-derived dendritic cells (DCs), a type of potent antigen-presenting cells. beta-glucan isolated from P. polymyxa JB115 enhanced the viability and induced the maturation of DCs. beta-glucan markedly increased the cytokine production of DCs and surface expression of DC markers. In addition, DCs treated with beta-glucan showed a higher capacity to stimulate allogeneic spleen cell proliferation compared to those treated with medium alone. These results demonstrate the effect of beta-glucan on DC maturation and may increase the use of beta-glucan.

Keyword

dendritic cells; JB115; maturation; Paenibacillus polymyxa; beta-glucan

MeSH Terms

Animals
Bone Marrow Cells/cytology/*drug effects/*immunology
Cell Survival/drug effects/*immunology
Dendritic Cells/cytology/*drug effects/*immunology
Flow Cytometry
Immunophenotyping/methods
Interleukin-12/analysis/immunology
Mice
Mice, Inbred BALB C
Nitric Oxide/analysis/immunology
Paenibacillus/*chemistry
Tumor Necrosis Factor-alpha/analysis/immunology
beta-Glucans/isolation & purification/*pharmacology

Figure

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  Fig. 1 Clusters of dendritic cells (DCs) treated with β-glucan. DCs were seeded at a density of 5 × 105 cells/mL in 6-well culture plates, and then treated with β-glucan for 2 days. (A) DC morphology. ×200. (B) The number and size of DCs was measured by flow cytometric analysis.

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  Fig. 2 Effect of β-glucan on DC viability. DCs were seeded at a concentration of 2.5 × 105 cells/mL in 96-well culture plates, and then treated with the indicated concentrations of β-glucan for 2 days. For the viability assay, treated cells were stained with a trypan blue staining solution to determine the number of live and dead cells. Data are presented as the mean ± SD from four individual wells. **p < 0.01.

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  Fig. 3 β-glucan increases cytokine and nitric oxide production of DCs. DCs were seeded and treated as described in Fig. 2. The amounts of interleukin (IL)-12 (A), tumor necrosis factor (TNF)-α (B), and nitric oxide (C) produced by DCs were measured. Data are presented as the mean ± SD from four individual wells. **p < 0.01, ***p < 0.001.

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  Fig. 4 β-glucan enhances the expression of immune-related DC surface markers but decreases DC antigen uptake. DCs were prepared and treated as described in the legend for Fig. 1. For surface marker analysis (A), the number in the histogram indicates the mean fluorescence intensity (MFI) of the main DC population. For antigen uptake analysis (B), DCs were incubated with 250 µg/mL dextran-fluorescein isothiocyanate at 4℃ or 37℃ for 45 min. The number in the histogram indicates the MFI of viable DCs.

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  Fig. 5 β-glucan increases the allo-proliferative capability of DCs. DC cultures were established and treated as described in the legend for Fig. 1. For the allo-proliferation assay, DCs treated with β-glucan (1 × 104 cells/well) were co-cultured for 5 days with allogeneic splenocytes (2 × 105 cells/well) stained with 5 µM carboxyfluorescein succinimidyl ester. The number indicates the percentage of proliferating cells with a low FL1 value in the co-culture.

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