Korean J Physiol Pharmacol.  2020 Jan;24(1):47-52. 10.4196/kjpp.2020.24.1.47.

Bordetella bronchiseptica is a potent and safe adjuvant that enhances the antigen-presenting capability of dendritic cells

• Lee YJ
• Han Y
• Joo HG

• KMID: 2466569
• DOI: http://doi.org/10.4196/kjpp.2020.24.1.47

Abstract

We previously demonstrated that Bordetella bronchiseptica (B. bronchiseptica) antigen (Ag) enhances the Mycoplasma hyopneumoniae Ag-specific immune response. The focus of this study was whether acellular bacterin of B. bronchiseptica could be used as an adjuvant to increase antigen-presenting capability of dendritic cells (DCs) by increasing the level of activation. The metabolic activity of DCs was increased by B. bronchiseptica, similar to lipopolysaccharide (LPS). Flow cytometry analysis revealed that B. bronchiseptica increases the expression of major histocompatibility complex class-2, cluster of differentiation (CD)40, CD54, and CD86 which are closely related to DC-mediated immune responses. B. bronchiseptica enhanced the production of cytokines related to adaptive immune responses. Furthermore, the survival rate of B. bronchiseptica-injected groups was 100% at 15 and 20 mg/kg doses, whereas that of LPS-injected groups was only 20%, 0% at 15 and 20 mg/kg doses respectively, and so B. bronchiseptica is likely to be safer than LPS. Taken together, these results indicate that B. bronchiseptica can be used as an adjuvant to enhance the antigen-presenting capability of DCs. B. bronchiseptica is a candidate for producing vaccines, especially in case of DC-mediating efficacy and safety demands. This study provides researchers and clinicians with valuable information regarding the usage of B. bronchiseptica as a safe bacteria-derived immunostimulating agent for developing efficient vaccines.

Keyword

Bordetella bronchiseptica; Dendritic cells; Immunostimulation; Safety; Vaccines

MeSH Terms

Bacterial Vaccines
Bordetella bronchiseptica*
Bordetella*
Cytokines
Dendritic Cells*
Flow Cytometry
Immunization
Major Histocompatibility Complex
Mycoplasma hyopneumoniae
Survival Rate
Vaccines
Bacterial Vaccines
Cytokines
Vaccines

Figure

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  Fig. 1 The effect of Bordetella bronchiseptica (B. bronchiseptica) on the metabolic activity of dendritic cells (DCs). DCs were cultured in 96-well plates at a concentration of 1.5 × 105 cells/ml with 0 to 300 ng/ml B. bronchiseptica or lipopolysaccharide (LPS) for 3 days. The treated DCs were incubated with 5 µl/well CCK-8 solution at 37℃ for 4 h. The optical density (O.D.) was measured at 450 nm by using a microplate reader. **Indicates p < 0.01.

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  Fig. 2 Effect of Bordetella bronchiseptica (B. bronchiseptica) on the expression of immune response-related surface markers of dendritic cells (DCs). DCs were cultured in 6-well plates at a concentration of 2.5 × 105 cells/ml and treated with 0.1 µg/ml B. bronchiseptica or lipopolysaccharide (LPS) for 2 days. After the treatment, DCs were stained as described in “Methods”. Numbers of histograms represent the geometric mean fluorescence intensity.

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  Fig. 3 Bordetella bronchiseptica (B. bronchiseptica) increases the level of dendritic cell (DC) maturation. DCs were cultured in 6-well plates at a concentration of 5.5 × 105 cells/ml and treated with 0.1 µg/ml of B. bronchiseptica or lipopolysaccharide (LPS) for 2 days. After treatment, the DCs were incubated with fluorescein isothiocyanate (FITC)-dextran as described in “Methods”. Numbers of histograms represent the geometric mean fluorescence intensity. Negative control DCs was treated at 4℃, and others were treated at 37℃ for 1 h.

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  Fig. 4 The effect of Bordetella bronchiseptica (B. bronchiseptica) on the production of immune-related cytokines. Dendritic cells (DCs) were cultured in 96-well plates at a concentration of 5 × 105 cells/ml and treated with 0 to 0.5 µg/ml B. bronchiseptica (BB) or lipopolysaccharide (LPS) for 3 days. The supernatant of cells treated with BB or LPS was collected and used for ELISA. The optical density was measured at 405 nm by using microplate reader. TNF, tumor necrosis factor; IL, interleukin. **,***Indicate p < 0.01, p < 0.001, respectively.

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  Fig. 5 Effect of Bordetella bronchiseptica (B. bronchiseptica) on the antigen-presenting capability of dendritic cells (DCs). DCs were treated with 0.1 µg/ml of LPS or B. bronchiseptica (BB) for 2 days. The treated DCs were incubated with 50 µg/ml mitomycin C for 30 min before the co-culture setup. Allogeneic spleen cells were harvested from Balb/c mice. DCs and allogeneic spleen cells were co-cultured in 96-well plates at a 1 × 105 cells/ml and 2 × 106 cells/ml respectively. After 5 days, CCK-8 assay was performed as described in ‘Methods’. O.D., optical density. **Indicates p < 0.01.

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  Fig. 6 Bordetella bronchiseptica (B. bronchiseptica) is safe in vivo. 7-week old Balb/c mice were intraperitoneally injected with B. bronchiseptica (BB) or lipopolysaccharide (LPS) at 10, 15, and 20 mg/kg dose respectively. The survival rate of the mice groups was determined at 7 day.

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