Brefeldin A and Cytochalasin B reduce dengue virus replication in cell cultures but do not protect mice against viral challenge

Farias, Kléber Juvenal Silva; Machado, Paula Renata Lima; Júnior, Renato Ferreira de Almeida; Fonseca, Benedito Antônio Lopes da

doi:10.1099/acmi.0.000041

Cited by 7 publications

(4 citation statements)

References 47 publications

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“…is could be explained by various factors. First, BFA has been reported to be an inhibitor of viral replication in the early stages of the viral cycle [46,47], which is consistent with the observations found here, where a decrease in the viral titer was observed only in cells treated with BFA after 1 h of infection (Figure 4(a)). On the other hand, NS1 secretion does not occur exclusively by the classical pathway, as reported by Alcalá et al, and treatment with BFA decreased NS1 secretion by only 35% [13].…”

Section: Discussionsupporting

confidence: 90%

Combination of the Focus-Forming Assay and Digital Automated Imaging Analysis for the Detection of Dengue and Zika Viral Loads in Cultures and Acute Disease

Bolívar-Marín

Bosch

Narváez

2022

Journal of Tropical Medicine

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Optimized methods for the detection of flavivirus infections in hyperendemic areas are still needed, especially for working with patient serum as a starting material. The focus-forming assay (FFA) reveals critical aspects of virus-host interactions, as it is a quantitative assay to determine viral loads. Automated image analysis provides evaluations of relative amounts of intracellular viral protein at the single-cell level. Here, we developed an optimized FFA for the detection of infectious Zika virus (ZIKV) and dengue virus (DENV) viral particles in cell cultures and clinical serum samples, respectively. Vero-76 cells were infected with DENV-2 (16681) or ZIKV (PRVA BC59). Using a panel of anti-DENV and anti-ZIKV NS1-specific monoclonal antibodies (mAbs), the primary mAbs, concentration, and the optimal time of infection were determined. To determine whether intracellular accumulation of NS1 improved the efficiency of the FFA, brefeldin A was added to the cultures. Focus formation was identified by conventional optical microscopy combined with CellProfiler™ automated image analysis software. The FFA was used with spike assays for ZIKV and clinical specimens from natural infection by DENV-1 and DENV-2. mAb 7744-644 for ZIKV and mAb 724-323 for DENV used at a concentration of 1 μg/ml and a time of 24 hours postinfection produced the best detection of foci when combining conventional counting and automated digital analysis. Brefeldin A did not improve the assessment of FFUs or their digitally assessed intensity at single-cell level. The FFA showed 95% ZIKV recovery and achieved the detection of circulating DENV-1 and DENV-2 in the plasma of acutely ill patients. The combination of the two techniques optimized the FFA, allowing the study of DENV and ZIKV in culture supernatants and clinical specimens from natural infection in hyperendemic areas.

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Section: Discussionsupporting

confidence: 90%

Combination of the Focus-Forming Assay and Digital Automated Imaging Analysis for the Detection of Dengue and Zika Viral Loads in Cultures and Acute Disease

Bolívar-Marín

Bosch

Narváez

2022

Journal of Tropical Medicine

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“…Brefeldin A is a fungal metabolite produced by species of the Ascomycetes [ 119 ], and to the best of our knowledge, it is being reported for the first time in bacterial endophytes. Brefeldin A has been reported to possess anticancer activity [ 101 ]; different cancer cell lines can be used to determine its anticancer activity with different cell lines.…”

Section: Resultsmentioning

confidence: 99%

Antibacterial and Anticancer Activity and Untargeted Secondary Metabolite Profiling of Crude Bacterial Endophyte Extracts from Crinum macowanii Baker Leaves

Sebola

Uche-Okereafor

Mekuto

et al. 2020

International Journal of Microbiology

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This study isolated and identified endophytic bacteria from the leaves of Crinum macowanii and investigated the potential of the bacterial endophyte extracts as antibacterial and anticancer agents and their subsequent secondary metabolites. Ethyl acetate extracts from the endophytes and the leaves (methanol: dichloromethane (1 : 1)) were used for antibacterial activity against selected pathogenic bacterial strains by using the broth microdilution method. The anticancer activity against the U87MG glioblastoma and A549 lung carcinoma cells was determined by the MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxy-phenyl)-2-(4-sulfophenyl)-2H-tetrazolium) assay. Bacterial endophytes that were successfully isolated from C. macowanii leaves include Raoultella ornithinolytica, Acinetobacter guillouiae, Pseudomonas sp., Pseudomonas palleroniana, Pseudomonas putida, Bacillus safensis, Enterobacter asburiae, Pseudomonas cichorii, and Arthrobacter pascens. Pseudomonas cichorii exhibited broad antibacterial activity against both Gram-negative and Gram-positive pathogenic bacteria while Arthrobacter pascens displayed the least MIC of 0.0625 mg/mL. Bacillus safensis crude extracts were the only sample that showed notable cell reduction of 50% against A549 lung carcinoma cells at a concentration of 100 μg/mL. Metabolite profiling of Bacillus safensis, Pseudomonas cichorii, and Arthrobacter pascens crude extracts revealed the presence of known antibacterial and/or anticancer agents such as lycorine (1), angustine (2), crinamidine (3), vasicinol (4), and powelline. It can be concluded that the crude bacterial endophyte extracts obtained from C. macowanii leaves can biosynthesize bioactive compounds and can be bioprospected for medical application into antibacterial and anticancer agents.

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“…Dengue is a systemic, febrile disease caused by a mosquito-borne virus known as Dengue virus (DENV) and is one of the major public health problems in tropical and subtropical countries [ 1 , 2 ]. DENV is a small (50nm) enveloped virus that belongs to the genus Flavivirus from family “Flaviviridae” [ 3 ]. Infection by any of the DENV serotype provides lifelong immunity against that particular serotype but only a few months of immunity against others.…”

Section: Introductionmentioning

confidence: 99%

A retrospective study on the socio-demographic factors and clinical parameters of dengue disease and their effects on the clinical course and recovery of the patients in a tertiary care hospital of Bangladesh

et al. 2022

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Dengue, a mosquito transmitted febrile viral disease, is a serious public health concern in Bangladesh. Despite significant number of incidences and reported deaths each year, there are inadequate number of studies relating the temporal trends of the clinical parameters as well as socio-demographic factors with the clinical course of the disease. Therefore, this study aims to associate the clinical parameters, demographic and behavioral factors of the dengue patients admitted in a tertiary care hospital in Dhaka, Bangladesh during the 2019 outbreak of dengue with the clinical course of the disease. Data were collected from the 336 confirmed dengue in-patients and analyzed using SPSS 26.0 software. Majority of the patients were male (2.2 times higher than female) who required longer time to recover compared to females (p < 0.01), urban resident (54.35%) and belonged to the age group of 18–40 years (73.33%). Dengue fever (90.77%) and dengue hemorrhagic fever (5.95%) were reported in most of the dengue patients while fever (98%) was the most frequently observed symptom. A significantly positive association was found between patient’s age and number of manifested symptoms (p = 0.013). Average duration of stay in the hospital was 4.9 days (SD = 1.652) and patient’s recovery time was positively correlated with delayed hospitalization (p < 0.01). Additionally, recovery time was negatively correlated with initial blood pressure (both systolic (p = 0.001, and diastolic (p = 0.023)) and platelet count (p = 0.003) of the patients recorded on the first day of hospitalization. Finally, a statistical model was developed which predicted that, hospital stay could be positively associated with an increasing trend of temperature, systolic blood pressure and reduced platelets count. Findings of this study may be beneficial to better understand the clinical course of the disease, identify the potential risk factors and ensure improved patient management during future dengue outbreaks.

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Brefeldin A and Cytochalasin B reduce dengue virus replication in cell cultures but do not protect mice against viral challenge

Cited by 7 publications

References 47 publications

Combination of the Focus-Forming Assay and Digital Automated Imaging Analysis for the Detection of Dengue and Zika Viral Loads in Cultures and Acute Disease

Combination of the Focus-Forming Assay and Digital Automated Imaging Analysis for the Detection of Dengue and Zika Viral Loads in Cultures and Acute Disease

Antibacterial and Anticancer Activity and Untargeted Secondary Metabolite Profiling of Crude Bacterial Endophyte Extracts from Crinum macowanii Baker Leaves

A retrospective study on the socio-demographic factors and clinical parameters of dengue disease and their effects on the clinical course and recovery of the patients in a tertiary care hospital of Bangladesh

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