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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,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>
( Alisha Wehdnesday Bernardo Reyes ),( Hannah Leah Tadeja Simborio ),( Huynhtanhop ),( Lauren Togonon Arayan ),( Won Gi Min ),( Hu Jang Lee ),( Hong Hee Chang ),( Suk Kim ) 한국예방수의학회 2014 예방수의학회지 Vol.38 No.4
Brucellosis is an important and re-emerging zoonotic disease worldwide. The prevention of human infection isachieved predominantly through the control of brucellosis in agricultural animals, which in turn depends on accurate diagnosisand vaccination. However, conventional serological diagnosis of brucellosis has several limitations, and currently availablevaccines for animals have several drawbacks, including the ability to cause infection in humans. Phosphoglycerate kinase(Pgk) is one of the specific proteins reactive with mouse sera in the early stage of Brucella infection, and deletion of thepgk gene in B. abortus strain 2308 resulted in extreme attenuation of this strain in vitro and in vivo. Furthermore, the B. abortus pgk mutant has been used as a live vaccine, and in challenge experiments, it induced protection that was superiorto that conferred by commercial strains. In this study, the pgk gene from Brucella abortus 544 was successfully amplified andcloned into a maltose binding protein fusion protein expression vector (pMAL). The recombinant protein was expressed inEscherichia coli DH5α and purified. The immunogenicity of purified recombinant B. abortus 544 Pgk (rPgk) was evaluatedby western blot analysis using Brucella-positive mouse sera. rPgk could be used as an antigenic component for futureserological tests and potential vaccine development.
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.
Reyes, Alisha Wehdnesday Bernardo,Hop, Huynh Tan,Arayan, Lauren Togonon,Huy, Tran Xuan Ngoc,Park, Soo Jong,Kim, Kwang Dong,Min, WonGi,Lee, Hu Jang,Rhee, Man Hee,Kwak, Yi-Seong,Kim, Suk Elsevier Ireland Ltd 2017 Journal of Ethnopharmacology Vol.198 No.-
<P><B>Abstract</B></P> <P><B>Ethnopharmacological relevance</B></P> <P> <I>Panax ginseng</I> Meyer (Araliaceae), is one of the most valuable traditional Chinese medicines and is used for the treatment of various human diseases. In this study, we elucidated the protective mechanism of the essential oil from Korean red ginseng (RGO) against <I>Brucella</I> infection.</P> <P><B>Materials and methods</B></P> <P>The effects of RGO on <I>Brucella abortus</I> viability, NO production, uptake and intracellular growth in macrophages were investigated. Mice were intraperitoneally infected with <I>B. abortus</I> and orally treated with RGO for 14 days. The weights and bacterial numbers from each spleen were monitored, and the sera were evaluated for cytokine production.</P> <P><B>Results</B></P> <P> <I>B. abortus</I> viability was not affected, whereas NO production, internalization and intracellular replication were inhibited in RGO-treated macrophages. Bacterial adherence, F-actin polymerization and MAPK signaling protein phosphorylation (ERK1/2, JNK and p38α) were reduced and the co-localization of <I>B. abortus-</I>containing phagosomes with LAMP-1 was augmented in RGO-treated cells compared to untreated cells. RGO displayed protective effects against cell damage by inhibiting nitrite production during <I>B. abortus</I> infection in macrophages. Moreover, the spleen weight and bacterial burden were lower in the RGO-treated group than in the control group. The uninfected RGO-treated mice displayed increased TNF-α and IFN-γ production, whereas the <I>B. abortus-</I>infected RGO-treated mice showed reduced IL-10 production compared to the control.</P> <P><B>Conclusion</B></P> <P>RGO exhibits protective effects against <I>B. abortus</I> infection <I>in vitro</I> and <I>in vivo,</I> which emphasize the beneficial effects of RGO in the prevention and treatment of brucellosis.</P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>
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.
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,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,Kim Heejin,Huy Tran Xuan Ngoc,Nguyen Trang Thi,Min Wongi,Lee Dongho,Hur Jin,Lee John Hwa,김석 한국미생물·생명공학회 2022 Journal of microbiology and biotechnology Vol.32 No.9
This study investigated the contribution of lipoxygenase (LOX) inhibitors, nordihydroguaiaretic acid (NDGA), tetra-O-methyl nordihydroguaiaretic acid (M4N) and zileuton (ZIL), and thromboxane A2 (TXA2) inhibitor 4,5-diphenylimidazole (DPI) in the proliferation of Brucella abortus infection. None of the compounds affected the uptake of Brucella into the macrophages. We determined the effect of neutralizing leukotriene B4 (LTB4) receptor and showed that the uptake of the bacteria was inhibited at 30 min post-infection. M4N treatment attenuated intracellular survival of Brucella at 2 h post-incubation but it was not observed in the succeeding time points. DPI treatment showed reduced survival of Brucella at 24 h post-incubation while blocking LTB4 receptor was observed to have a lower intracellular growth at 48 h post-incubation suggesting different action of the inhibitors in the course of the survival of Brucella within the cells. Reduced proliferation of the bacteria in the spleens of mice was observed in animals treated with ZIL or DPI. Increased serum cytokine level of TNF-α and MCP-1 was observed in mice treated with M4N or ZIL while a lower IFN-γ level in ZIL-treated mice and a higher IL-12 serum level in DPI-treated mice were observed at 7 d postinfection. At 14 d post-infection, ZIL-treated mice displayed reduced serum level of IL-12 and IL-10. Overall, inhibition of 5-LOX or TXA2 or a combination therapy promises a potential alternative therapy against B. abortus infection. Furthermore, strong ligands for LTB4 receptor could also be a good candidate for the control of Brucella infection.
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.