J Vet Sci.  2018 Jan;19(1):51-57. 10.4142/jvs.2018.19.1.51.

Tannic acid-mediated immune activation attenuates Brucella abortus infection in mice

• Reyes AW
• Hop HT
• Arayan LT
• Huy TX
• Min W
• Lee HJ
• Chang HH
• Kim S

• KMID: 2402695
• DOI: http://doi.org/10.4142/jvs.2018.19.1.51

Abstract

Brucellosis is an emerging infectious disease affecting humans and animals. In this study, we investigated the in vitro and in vivo effects of tannic acid (TA) against Brucella abortus infection. After infection, F-actin polymerization and mitogen-activated protein kinases (MAPKs) (ERK 1/2 and p38α) phosphorylation were reduced in TA-treated cells compared with that in control cells. The mice were infected via an intraperitoneal route and were orally given TA or phosphate-buffered saline for 14 days. Spleen weights of the TA-treated and control mice were not different; however, splenic proliferation of B. abortus was significantly reduced in the TA-treated group. Immune response analysis showed that, compared with the control group, non-infected TA-treated mice displayed increased levels of interferon-γ (IFN-γ), monocyte chemoattractant protein-1 (MCP-1), and interleukin-10 at 3 days post-infection and a further increase in IFN-γ and MCP-1 at 14 days post-infection. In contrast, compared with the control group, infected TA-treated mice displayed elevated levels of IFN-γ at 3 days post-infection, which continued to increase at 14 days post-infection, as was also observed for tumor necrosis factor. Taken together, the results showing TA activation of cytokine production and inhibition of bacterial proliferation in the host highlight a potential use of TA treatment in the control of Brucella infection.

Keyword

Brucella abortus; actins; cytokines; mitogen-activated protein kinase; tannins

MeSH Terms

Actins
Animals
Brucella abortus*
Brucella*
Brucellosis
Chemokine CCL2
Communicable Diseases, Emerging
Cytokines
Humans
In Vitro Techniques
Interleukin-10
Mice*
Mitogen-Activated Protein Kinases
Phosphorylation
Polymerization
Polymers
Spleen
Tannins
Tumor Necrosis Factor-alpha
Weights and Measures
Actins
Chemokine CCL2
Cytokines
Interleukin-10
Mitogen-Activated Protein Kinases
Polymers
Tannins
Tumor Necrosis Factor-alpha

Figure

• 

  Fig. 1 Effect of tannic acid (TA) on F-actin polymerization in RAW 264.7 cells. (A) The stained cells were viewed under a laser scanning confocal microscope. (B) F-actin content was quantified using a flow cytometer. The values expressed are means ± SD for each group. A statistically significant difference from the control group is indicated by an asterisk (*p < 0.05). DIC, differential interference contrast; B. abortus, Brucella abortus.

• 

  Fig. 2 Effect of tannic acid (TA) on mitogen-activated protein kinase (MAPK) phosphorylation in RAW 264.7 cells. Cells were pre-treated with TA or PBS. The cells were then infected with Brucella abortus and the cell lysates collected. Immunoblot analysis of total RAW 264.7 cell lysates was assessed by using phospho-specific ERK1/2, JNK, and p38α antibodies.

• 

  Fig. 3 Total weight of spleens of mice orally treated with tannic acid (TA) or PBS. Mice were orally given TA or PBS from 3 days prior to infection to 14 days post-infection. The mice were then sacrificed, the spleens harvested and weighed. A statistically significant difference from the control group is indicated by an asterisk (*p < 0.05).

• 

  Fig. 4 Proliferation of Brucella abortus in spleens of tannic acid (TA)-treated mice or PBS-treated mice. Mice were orally given TA or PBS from 3 days prior to infection to 14 days post-infection. At 14 days post-infection, mice were sacrificed, and the spleens homogenized and serially diluted in PBS. The number of colony-forming units (CFUs) was counted to assess bacterial proliferation in each spleen. A statistically significant difference from the control group is indicated by an asterisk (**p < 0.01).

• 

  Fig. 5 Immune response analysis of tannic acid (TA)-treated mice or PBS-treated mice against Brucella abortus infection in mice. Mice were orally given TA or PBS from 3 days prior to infection to 14 days post-infection. At 3 days post-infection, serum samples were collected via tail vein in each animal. At 14 days post-infection, mice were sacrificed, and blood was collected from the heart. Serum samples were analyzed for cytokine release by using a flow cytometer. A statistically significant difference from the control group is indicated by asterisks (*p < 0.05; **p < 0.01; ***p < 0.001). IL, interleukin; MCP-1, monocyte chemoattractant protein-1; TNF, tumor necrosis factor; IFN, interferon.

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