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( Alisha Wehdnesday Bernardo Reyes ),( Hannah Leah Tadeja Simborio ),( Huynh Tan Hop ),( Lauren Togonon Arayan ),( Tran Xuan Ngoc Huy ),( Wongi Min ),( Suk Kim ) 한국예방수의학회(구 한국수의공중보건학회) 2015 예방수의학회지 Vol.39 No.4
The bacterial lipopolysaccharide (LPS) mainly contributes to the structural integrity, survival and protection barrier against harsh environments. Therefore, the early stages in LPS or lipid A biosynthesis are attractive targets in the identification and development of inhibitors which would be effective against infections caused by Gram-negative bacteria. The bacterial outer membrane proteins (OMPs) meanwhile function as maintenance for structure, adhesion to other cells and substances, as well as development of resistance to antimicrobials. The LPS and LPS-related molecules, and OMPs are important immunogenic components of several important pathogens including Brucella, which have been extensively used in immunological studies and in the diagnosis of diseases. Here we review the importance, structure, functions and immunogenic aspects of LPS and OMPs particularly of Brucella which can be targeted for the prevention and diagnosis of brucellosis.
Reyes, Alisha Wehdnesday Bernardo,Hong, Tae Gyu,Hop, Huynh Tan,Arayan, Lauren Togonon,Huy, Tran Xuan Ngoc,Min, Wongi,Lee, Hu Jang,Lee, Kang Seok,Kim, Suk Elsevier 2017 Microbial pathogenesis Vol.109 No.-
<P><B>Abstract</B></P> <P>In this study, we investigated the protective effects of tannin-derived components, gallic acid (GA) and tannic acid (TA), <I>in vitro</I> and <I>in vivo</I> against <I>Salmonella</I> infection in mice. Both GA and TA showed antibacterial effects against <I>Salmonella</I> (<I>S.</I>) Typhimurium as well as inhibitory effects on the adherence, invasion, and intracellular growth of the pathogens in macrophages. Following a lethal dose of <I>Salmonella</I> infection in mice, reduced virulence in both GA- and TA-treated groups was observed based on reduced mortality rates. In the non-infected groups, the average weights of the spleens and livers of GA- or TA-treated mice were not significantly different with the control group. In addition, the average weights of these organs in all of the <I>Salmonella</I>-infected groups were not significantly different but the numbers of bacteria in the spleens and livers in both GA- and TA-treated mice were significantly reduced. The levels of cytokine production in non-infected mice revealed that GA-treated and TA-treated mice elicited an increased level of IFN-γ, and both IFN-γ and MCP-1, respectively, as compared with the PBS-treated group. These findings highlight the potential of GA and TA as alternatives for the treatment of salmonellosis and as supplements to conventional antimicrobial food additives.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Gallic acid (GA) and tannic acid (TA) showed antibacterial effects against <I>S.</I> Typhimurium. </LI> <LI> GA and TA showed a treatment effect against <I>S.</I> Typhimurium infection in a mouse model. </LI> <LI> GA-treated and TA-treated mice elicited an increased level of IFN-γ, and both IFN-γ and MCP-1, respectively. </LI> <LI> GA and TA could be used as potential alternatives for the treatment of salmonellosis. </LI> </UL> </P>
Reyes, Alisha Wehdnesday Bernardo,Arayan, Lauren Togonon,Huy, Tran Xuan Ngoc,Vu, Son Hai,Kang, Chang Keun,Min, Wongi,Lee, Hu Jang,Lee, John Hwa,Kim, Suk Elsevier 2019 Veterinary microbiology Vol.237 No.-
<P><B>Abstract</B></P> <P>We investigated the involvement of chemokine receptor type 4 (CXCR4) signaling on the outcome of <I>Brucella</I> (<I>B.</I>) <I>abortus</I> 544 infection in murine macrophages and in a mouse model. CXCR4 manipulation were first evaluated for <I>Brucella</I> invasion and intracellular survival efficiency, mitogen-activated protein kinases (ERK1/2, JNK, p38α) activation and generation of nitric oxide (NO), and then in the splenic bacterial proliferation and cytokine production in BALB/c mice. CXCR4 blockade is involved in the successful control of <I>Brucella</I> invasion, reduction of ERK1/2 phosphorylation and inhibition of nitric oxide release from macrophages. Furthermore, using a reported CXCR4-specific antagonist AMD3100 resulted in splenomegaly but attenuated <I>Brucella</I> proliferation in these organs with elevated serum levels of MCP-1, TNF and IL-12. These findings provide insights on the contribution of CXCR4 signaling in the phagocytic pathway and immune modulation during <I>B. abortus</I> infection.</P> <P><B>Highlights</B></P> <P> <UL> <LI> CXCR4 blockade attenuated <I>B. abortus</I> invasion into RAW264.7 cells possibly via ERK pathway. </LI> <LI> CXCR4 blockade inhibited generation of NO in RAW264.7 cells during <I>B. abortus</I> infection. </LI> <LI> AMD3100 treatment in mice did not negatively affect body weight, serum ALT concentration or weight of spleens. </LI> <LI> AMD3100 treatment in mice displayed splenomegaly but reduced <I>B. abortus</I> proliferation with elevated serum levels of MCP-1, TNF and IL-12. </LI> </UL> </P>
Reyes Alisha Wehdnesday Bernardo,Kim Heejin,Huy Tran Xuan Ngoc,Nguyen Trang Thi,Min Wongi,Lee Hu Jang,Hur Jin,Lee John Hwa,Kim Suk 한국미생물·생명공학회 2023 Journal of microbiology and biotechnology Vol.33 No.4
Brucellosis is a contagious zoonotic disease that infects millions of people annually with hundreds of millions more being exposed. It is caused by Brucella, a highly infectious bacterial species capable of infecting humans with an estimated dose of 10-100 organisms. Sirtuin 1 (SIRT1) has been reported to contribute to prevention of viral diseases as well as a chronic infection caused by Mycobacterium bovis. Here, we investigated the role of SIRT1 in the establishment of Brucella abortus infection in both in vitro and in vivo systems using the reported SIRT1 activators resveratrol (RES), piceatannol (PIC), and ginsenoside Rg3 (Rg3). In RAW264.7 cells, SIRT1 activators did not alter the adherence of Brucella or Salmonella Typhimurium. However, reduced uptake of Brucella was observed in cells treated with PIC and Rg3, and survival of Brucella within the cells was only observed to decrease in cells that were treated with Rg3, while PIC treatment reduced the intracellular survival of Salmonella. SIRT1 treatment in mice via oral route resulted in augmented Brucella resistance for PIC and Rg3, but not RES. PIC treatment favors Th2 immune response despite reduced serum proinflammatory cytokine production, while Rg3-treated mice displayed high IL-12 and IFN-γ serum production. Overall, our findings encourage further investigation into the complete mechanisms of action of the different SIRT1 activators used as well as their potential benefit as an effective alternative approach against intracellular and extracellular pathogens.
Alisha Wehdnesday Bernardo Reyes 대한수의학회 2021 Journal of Veterinary Science Vol.22 No.2
Background: We previously elucidated the protective mechanism of Korean red ginseng oil (RGO) against Brucella abortus infection, and our phytochemical analysis revealed that palmitic acid (PA) was an abundant component of RGO. Consequently, we investigated the contribution of PA against B. abortus. Objectives: We aimed to investigate the efficacy of PA against B. abortus infection using a murine cell line and a murine model. Methods: Cell viability, bactericidal, internalization, and intracellular replication, western blot, nitric oxide (NO), and superoxide (O2-) analyses and flow cytometry were performed to determine the effects of PA on the progression of B. abortus infection in macrophages. Flow cytometry for cytokine analysis of serum samples and bacterial counts from the spleens were performed to determine the effect of PA in a mouse model. Results: PA did not affect the growth of B. abortus. PA treatment in macrophages did not change B. abortus uptake but it did attenuate the intracellular survivability of B. abortus. Incubation of cells with PA resulted in a modest increase in sirtuin 1 (SIRT1) expression. Compared to control cells, reduced nitrite accumulation, augmented O2-, and enhanced pro-inflammatory cytokine production were observed in PA-treated B. abortus-infected cells. Mice orally treated with PA displayed a decreased serum interleukin-10 level and enhanced bacterial resistance. Conclusions: Our results suggest that PA participates in the control of B. abortus within murine macrophages, and the in vivo study results confirm its efficacy against the infection. However, further investigations are encouraged to completely characterize the mechanisms involved in the inhibition of B. abortus infection by fatty acids.
Alisha Wehdnesday Bernardo Reyes,Lauren Togonon Arayan,Huynh Tan Hop,Tran Xuan Ngoc Huy,Vu Hai Son,Suk Kim 한국예방수의학회 2018 예방수의학회지 Vol.42 No.2
We investigated the effects of two Brucella proteins expressed in a pMAL expression system, RocF and EF-Ts, as subunit vaccines on immune modulation and protective efficacy using a mouse model. Mice vaccinated with MBP-RocF and MBP-EF-Ts displayed increased production of TNF, IFN-r, MCP-1, IL-10 and IL-6, and TNF and MCP-1, respectively. Furthermore, mice vaccinated with MBP-EF-Ts showed decreased induction of IFN-r and Th2-related cytokines, IL-10 and IL-6. Higher proportions of CD4+ and CD8+ T cells were observed in the blood of mice vaccinated with MBP-RocF than in the PBS-vaccinated group, although the increases were not significant. Furthermore, significantly reduced Brucella proliferation in the spleens of the MBP-RocF and MBP-EF-Ts groups were observed, but inflammation of these organs was not attenuated. Overall, these results indicate that RocF and EF-Ts could be potential subunit vaccine candidates against animal brucellosis.
Alisha Wehdnesday Bernardo Reyes,김희진(Heejin Kim),Tran Xuan Ngoc Huy,Trang Thi Nguyen,민원기(Wongi Min),김현진Hyun Jin Kim),이후장(Hu Jang Lee),김석(Suk Kim) 한국예방수의학회 2021 예방수의학회지 Vol.45 No.4
We investigated the effect of a synthetic complement peptide C3a on the outcome of Brucella abortus 544 infection in a murine macrophage cell line RAW264.7 cell. First, we determined the highest non-cytotoxic concentration of the peptide in the cell line. We also found that the peptide significantly increased the growth of the bacteria at 8 and 24 h. Although the number of bacterial CFU was also elevated at 48 and 72 h, the increases were not significant as compared to controls. We further investigated the effect of C3a peptide on the growth of Brucella by pre-incubating the peptide at various temperatures and found that the effect was reversed at 24 h post-incubation suggesting that incubation of peptide at high temperatures including 65°C or 95°C could inactivate its action. This also could indicate the beneficial effect of high temperature during infection. Although several studies reported the inhibitory effect of different antimicrobial peptides including C3a, the present study preliminarily revealed that it had no positive contribution on the control of B. abortus 544 infection in vitro and indirectly to its receptor, CD88, which belongs to GPCR. Moreover, the encouraged further exploration of the effect of other similar peptides would be performed for the purpose of finding Brucella-host cell interaction for the control of disease progression.
Alisha Wehdnesday Bernardo Reyes,Lauren Togonon Arayan,TRAN XUAN NGOC HUY,Son Hai Vu,민원기,허진,김석 한국미생물·생명공학회 2020 Journal of microbiology and biotechnology Vol.30 No.4
We previously identified β-sitosterol (BS) as one of the most abundant compounds found in Korean red ginseng oil. BS is a widely prevalent vegetable-derived phytosterol with many known health benefits. Here, we investigated the efficacy of BS against Brucella (B.) abortus infection. BS showed no effect on bacterial growth but attenuated internalization, intracellular survival and MAPKslinked intracellular signaling in RAW264.7 cells. BS treatment in cells is also associated with increased nitrite concentration during infection at 24 h. Slightly enhanced resistance to B. abortus infection was observed in mice orally given BS, which could be mediated by induced production of proinflammatory cytokines. Taken together, our study demonstrates the contribution of BS treatment against B. abortus infection although further investigation is encouraged to maximize its beneficial effects against intracellular infection.
Alisha Wehdnesday Bernardo Reyes,Son Hai Vu,TRAN XUAN NGOC HUY,민원기,이후장,장홍희,이존화,김석 한국미생물·생명공학회 2020 Journal of microbiology and biotechnology Vol.30 No.5
In this study, we investigated the effects of linoleic acid (LA) treatment on Brucella abortus infection in professional phagocyte RAW264.7 cells, particularly during the pathogen’s invasion and intracellular growth in these cells, as well as in murine model BALB/c mice focusing on bacterial splenic proliferation and immunoregulatory activities. LA inhibited the growth of Brucella in a doseand time-dependent manner. The ability of the pathogen to enter the phagocytes was inhibited as was its survival within these cells. This was accompanied by increased nitrite accumulation in these cells at 24 h post-infection. The concentration of LA used in the present study did not affect the total body weight or liver function of the mice. During Brucella infection, the total splenic weight of these animals was not changed; rather, resistance to bacterial proliferation was enhanced in the spleen. Furthermore, mice treated with LA displayed elevated levels of IL-12 and IFN-γ but reduced levels of IL-10 during infection. The findings in this study showed the regulatory role of LA against B. abortus infection suggesting its potential use in designing intervention strategy for brucellosis.
Reyes, Alisha Wehdnesday Bernardo,Arayan, Lauren Togonon,Hop, Huynh Tan,Ngoc Huy, Tran Xuan,Vu, Son Hai,Min, WonGi,Lee, Hu Jang,Kim, Suk Elsevier 2018 Microbial pathogenesis Vol.119 No.-
<P><B>Abstract</B></P> <P>In this study, we investigated the effects of gallic acid (GA) in intracellular signaling within murine macrophages and its contribution to host immunity during <I>Brucella</I> infection<I>. In vitro</I> analysis revealed that GA treatment decreased F-actin content and suppressed p38α phosphorylation level. <I>In vivo</I> analysis showed that GA treatment reduced inflammation and proliferation of <I>Brucella</I> in spleens of mice in comparison to PBS treatment yielding a significant protection unit. For the analysis of immune response, the uninfected GA-treated mice showed increased production of IFN-γ and MCP-1, and the <I>Brucella-</I>infected GA-treated mice showed elevated levels of IL-12p70, TNF, IFN-γ, MCP-1, IL-10 and IL-6 in comparison to negative and positive control groups, respectively. These findings demonstrate the therapeutic effects of GA against <I>Brucella</I> infection through interference on intracellular signaling pathway, induction of cytokine production and protection from bacterial proliferation in spleens of mice.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Gallic acid (GA) decreased F-actin content and p38α phosphorylation level in RAW 264.7 cells. </LI> <LI> GA reduced inflammation and proliferation of <I>Brucella</I> in the spleens of mice. </LI> <LI> GA-treated mice showed increased production of IFN-γ and MCP-1. </LI> <LI> <I>Brucella</I>-infected GA-treated mice showed elevated levels of IL-12p70, TNF, IFN-γ, MCP-1, IL-10 and IL-6. </LI> <LI> GA treatment revealed potential therapeutic effects against <I>Brucella</I> infection in animals. </LI> </UL> </P>