Alisha Reyes - Academia.edu (original) (raw)
Papers by Alisha Reyes
Table S1a. Effect of PAFR activation on B. abortus internalization into macrophages. Table S1b. E... more Table S1a. Effect of PAFR activation on B. abortus internalization into macrophages. Table S1b. Effect of PAFR activation with different infection doses of B. abortus. (DOCX 22 kb)
Role of PAFR activation on the intensification of F-actin polymerization for phagocytosis of B. a... more Role of PAFR activation on the intensification of F-actin polymerization for phagocytosis of B. abortus. RAW 264.7 cells were pretreated for 1 h with a CV3988 (1 μM), followed by infection with B. abortus for 5 (a), 15 (b), or 30 (c) min, and then cells were subjected to the FACS analysis for F-actin content. (d) The quantitative analysis results of experiment in (a-c). Data represent the mean ± SD of triplicate trials from three independent experiments. Statistically significant differences from the untreated samples are indicated by asterisks (*, P <0.05, **, P
Uptake of B. abortus facilitates the internalization of PAFR from membrane. (a) RAW 264.7 cells w... more Uptake of B. abortus facilitates the internalization of PAFR from membrane. (a) RAW 264.7 cells were pretreated for 5 min with a PAF (200 nM), followed by infection with B. abortus for indicated times. Cell plasma membranes were isolated and monitored for the immunoblot analysis using an antibody against PAFR. Sodium potassium ATPase (Na/K ATPase) antibody was applied for quality control of plasma membrane. Images shown are representative of three independent experiments. (b) PAFR levels were quantified by fold activation detected as standardized ratio of PAFR to β-actin over basal levels present in resting cells. Data represent the mean ± SD of triplicate trials from three independent experiments. Statistically significant differences from the untreated samples are indicated by asterisks (**, P <0.01). (TIF 144 kb)
PAFR activation-related F-actin polymerization for phagocytosis of B. abortus with different infe... more PAFR activation-related F-actin polymerization for phagocytosis of B. abortus with different infection doses. RAW 264.7 cells were pretreated with or without PAF (200 nM) for 5 min, followed by infection with B. abortus (MOI 10, 50 and 100) for 5 (a) and 30 (b) min, and then cells were subjected to the FACS analysis for F-actin content. (c) The quantitative analysis results of experiment in (a-b). Data represent the mean ± SD of triplicate trials from three independent experiments. Statistically significant differences from the infected cells with B. abortus (MOI 10) are indicated by asterisks (*, P <0.05, **, P
To date, most serodiagnostic methods for brucellosis screening are based on antibodies against li... more To date, most serodiagnostic methods for brucellosis screening are based on antibodies against lipopolysaccharides of Brucella spp. However, this approach has the drawback of yielding false-positive results due to cross-reactivity with lipopolysaccharides of other related pathogens, especially Yersinia enterocolitica O:9. In this study, Brucella abortus AspC was cloned and expressed by PCR amplification into a pCold TF expression system to obtain recombinant AspC (rAspC). The immunogenicity of rAspC was confirmed by western blotting of Brucella-positive bovine serum. rAspC-based ELISA was performed to determine whether rAspC could be used in the serodiagnosis of bovine brucellosis. rAspC reacted strongly with anti-Brucella antibodies in positive sera in the tube agglutination test (TAT), but did not show strong reaction with most negative samples. In particular, the average OD492 value at the highest TAT titer showed a 1.4-fold increase with respect to the cutoff value. The accuracy...
Journal of the Preventive Veterinary Medicine, 2019
Veterinary Microbiology, 2020
Brucella as a stealthy intracellular pathogen avoids activation of innate immune response. Here w... more Brucella as a stealthy intracellular pathogen avoids activation of innate immune response. Here we investigated the contribution of an adenosine receptor, Adora2b, during Brucella infection in professional phagocyte RAW 264.7 cells and in a murine model. Adora2b-deficient cells showed attenuated Brucella internalization and intracellular survival with enhanced release of IL-6, TNF-α, IL-12 and MCP-1. In addition, blockade of Adora2b using MRS 1754 treatment in mice resulted in increased total weight of the spleens but suppressed bacterial burden in these organs accompanied by elevated levels of IL-6, IFN-γ, TNF-α, IL-12 and MCP-1, while reduced IL-10. Overall, we proposed that the Adora2b participates in the successful phagocytic pathway and intracellular survival of Brucella in RAW 264.7 cells, and could be a potential therapeutic target for the treatment of acute brucellosis in animals.
Journal of the Preventive Veterinary Medicine, 2017
Journal of the Preventive Veterinary Medicine, 2017
Frontiers in cellular and infection microbiology, 2018
The cellular oncogene c-Fos (c-Fos) is a component of activator protein 1 (AP1), a master transcr... more The cellular oncogene c-Fos (c-Fos) is a component of activator protein 1 (AP1), a master transcriptional regulator of cells. The suppression of c-Fos signaling by siRNA treatment resulted in significant induction of TLR4, which subsequently activates p38 and ERK1/2 mitogen-activated protein kinases (MAPKs) and enhances F-actin polymerization, leading to an increase in phagocytosis. During infection, c-Fos signaling is induced, which activates the downstream innate-immunity signaling cascade for bacterial clearance. The inhibition of c-Fos signaling led to increased production of interleukin 10 (IL-10), which partially suppressed lysosome-mediated killing, resulting in increased survival of inside macrophages. We present evidence of the regulatory role played by the c-Fos pathway in proliferation during infection; however, this was independent of the anti- effect of this pathway. Another finding is the essential contribution of c-Fos/TRAIL to infected-cell necrosis, which is a key e...
Journal of microbiology and biotechnology, Jan 3, 2018
The aim of this work is to investigate the protective efficacy of emodin, an active naturally occ... more The aim of this work is to investigate the protective efficacy of emodin, an active naturally occurring anthraquinone derivative of several traditional Chinese herbs, against infection in macrophages. were incubated with different concentrations of emodin and showed that bacterial survival rates were markedly reduced in a dose-dependent manner at increasing incubation time points. Through bacterial infection assay, the highest non-cytotoxic concentration of emodin demonstrated attenuated invasion of into macrophages, however it did not inhibit the growth of these pathogens within the host cells. On the other hand, emodin effectively decreased the number of bacteria that adhered to host cells, which indicated its potential as an anti-adhesin agent. Furthermore, using immunoblotting and FACS assay for detecting MAPK signaling proteins and F-actin polymerization, respectively, the results showed that the emodin-incubated cells displayed modest reduction in the phosphorylation levels of...
FEMS Microbiology Letters, 2015
This study highlights the phagocytic and intracellular modulating effect of RGSF-A and its potent... more This study highlights the phagocytic and intracellular modulating effect of RGSF-A and its potential as an alternative remedy to control Brucella abortus infection.
Journal of Veterinary Science, 2021
Journal of Microbiology and Biotechnology
Journal of the Preventive Veterinary Medicine
Journal of Microbiology and Biotechnology
Journal of Agriculture & Life Science
Journal of the Preventive Veterinary Medicine
Journal of the Preventive Veterinary Medicine
Table S1a. Effect of PAFR activation on B. abortus internalization into macrophages. Table S1b. E... more Table S1a. Effect of PAFR activation on B. abortus internalization into macrophages. Table S1b. Effect of PAFR activation with different infection doses of B. abortus. (DOCX 22 kb)
Role of PAFR activation on the intensification of F-actin polymerization for phagocytosis of B. a... more Role of PAFR activation on the intensification of F-actin polymerization for phagocytosis of B. abortus. RAW 264.7 cells were pretreated for 1 h with a CV3988 (1 μM), followed by infection with B. abortus for 5 (a), 15 (b), or 30 (c) min, and then cells were subjected to the FACS analysis for F-actin content. (d) The quantitative analysis results of experiment in (a-c). Data represent the mean ± SD of triplicate trials from three independent experiments. Statistically significant differences from the untreated samples are indicated by asterisks (*, P <0.05, **, P
Uptake of B. abortus facilitates the internalization of PAFR from membrane. (a) RAW 264.7 cells w... more Uptake of B. abortus facilitates the internalization of PAFR from membrane. (a) RAW 264.7 cells were pretreated for 5 min with a PAF (200 nM), followed by infection with B. abortus for indicated times. Cell plasma membranes were isolated and monitored for the immunoblot analysis using an antibody against PAFR. Sodium potassium ATPase (Na/K ATPase) antibody was applied for quality control of plasma membrane. Images shown are representative of three independent experiments. (b) PAFR levels were quantified by fold activation detected as standardized ratio of PAFR to β-actin over basal levels present in resting cells. Data represent the mean ± SD of triplicate trials from three independent experiments. Statistically significant differences from the untreated samples are indicated by asterisks (**, P <0.01). (TIF 144 kb)
PAFR activation-related F-actin polymerization for phagocytosis of B. abortus with different infe... more PAFR activation-related F-actin polymerization for phagocytosis of B. abortus with different infection doses. RAW 264.7 cells were pretreated with or without PAF (200 nM) for 5 min, followed by infection with B. abortus (MOI 10, 50 and 100) for 5 (a) and 30 (b) min, and then cells were subjected to the FACS analysis for F-actin content. (c) The quantitative analysis results of experiment in (a-b). Data represent the mean ± SD of triplicate trials from three independent experiments. Statistically significant differences from the infected cells with B. abortus (MOI 10) are indicated by asterisks (*, P <0.05, **, P
To date, most serodiagnostic methods for brucellosis screening are based on antibodies against li... more To date, most serodiagnostic methods for brucellosis screening are based on antibodies against lipopolysaccharides of Brucella spp. However, this approach has the drawback of yielding false-positive results due to cross-reactivity with lipopolysaccharides of other related pathogens, especially Yersinia enterocolitica O:9. In this study, Brucella abortus AspC was cloned and expressed by PCR amplification into a pCold TF expression system to obtain recombinant AspC (rAspC). The immunogenicity of rAspC was confirmed by western blotting of Brucella-positive bovine serum. rAspC-based ELISA was performed to determine whether rAspC could be used in the serodiagnosis of bovine brucellosis. rAspC reacted strongly with anti-Brucella antibodies in positive sera in the tube agglutination test (TAT), but did not show strong reaction with most negative samples. In particular, the average OD492 value at the highest TAT titer showed a 1.4-fold increase with respect to the cutoff value. The accuracy...
Journal of the Preventive Veterinary Medicine, 2019
Veterinary Microbiology, 2020
Brucella as a stealthy intracellular pathogen avoids activation of innate immune response. Here w... more Brucella as a stealthy intracellular pathogen avoids activation of innate immune response. Here we investigated the contribution of an adenosine receptor, Adora2b, during Brucella infection in professional phagocyte RAW 264.7 cells and in a murine model. Adora2b-deficient cells showed attenuated Brucella internalization and intracellular survival with enhanced release of IL-6, TNF-α, IL-12 and MCP-1. In addition, blockade of Adora2b using MRS 1754 treatment in mice resulted in increased total weight of the spleens but suppressed bacterial burden in these organs accompanied by elevated levels of IL-6, IFN-γ, TNF-α, IL-12 and MCP-1, while reduced IL-10. Overall, we proposed that the Adora2b participates in the successful phagocytic pathway and intracellular survival of Brucella in RAW 264.7 cells, and could be a potential therapeutic target for the treatment of acute brucellosis in animals.
Journal of the Preventive Veterinary Medicine, 2017
Journal of the Preventive Veterinary Medicine, 2017
Frontiers in cellular and infection microbiology, 2018
The cellular oncogene c-Fos (c-Fos) is a component of activator protein 1 (AP1), a master transcr... more The cellular oncogene c-Fos (c-Fos) is a component of activator protein 1 (AP1), a master transcriptional regulator of cells. The suppression of c-Fos signaling by siRNA treatment resulted in significant induction of TLR4, which subsequently activates p38 and ERK1/2 mitogen-activated protein kinases (MAPKs) and enhances F-actin polymerization, leading to an increase in phagocytosis. During infection, c-Fos signaling is induced, which activates the downstream innate-immunity signaling cascade for bacterial clearance. The inhibition of c-Fos signaling led to increased production of interleukin 10 (IL-10), which partially suppressed lysosome-mediated killing, resulting in increased survival of inside macrophages. We present evidence of the regulatory role played by the c-Fos pathway in proliferation during infection; however, this was independent of the anti- effect of this pathway. Another finding is the essential contribution of c-Fos/TRAIL to infected-cell necrosis, which is a key e...
Journal of microbiology and biotechnology, Jan 3, 2018
The aim of this work is to investigate the protective efficacy of emodin, an active naturally occ... more The aim of this work is to investigate the protective efficacy of emodin, an active naturally occurring anthraquinone derivative of several traditional Chinese herbs, against infection in macrophages. were incubated with different concentrations of emodin and showed that bacterial survival rates were markedly reduced in a dose-dependent manner at increasing incubation time points. Through bacterial infection assay, the highest non-cytotoxic concentration of emodin demonstrated attenuated invasion of into macrophages, however it did not inhibit the growth of these pathogens within the host cells. On the other hand, emodin effectively decreased the number of bacteria that adhered to host cells, which indicated its potential as an anti-adhesin agent. Furthermore, using immunoblotting and FACS assay for detecting MAPK signaling proteins and F-actin polymerization, respectively, the results showed that the emodin-incubated cells displayed modest reduction in the phosphorylation levels of...
FEMS Microbiology Letters, 2015
This study highlights the phagocytic and intracellular modulating effect of RGSF-A and its potent... more This study highlights the phagocytic and intracellular modulating effect of RGSF-A and its potential as an alternative remedy to control Brucella abortus infection.
Journal of Veterinary Science, 2021
Journal of Microbiology and Biotechnology
Journal of the Preventive Veterinary Medicine
Journal of Microbiology and Biotechnology
Journal of Agriculture & Life Science
Journal of the Preventive Veterinary Medicine
Journal of the Preventive Veterinary Medicine