Development of dengue virus plaques under serum-free overlay medium

Malewicz, Barbara; Jenkin, Howard M.

doi:10.1128/jcm.9.5.609-614.1979

Cited by 19 publications

(4 citation statements)

References 21 publications

(26 reference statements)

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“…In this study, we used Vero and BHK-21 cells as they are susceptible to DENV infection. In previous in vitro studies, various host cells, including BHK-2 16 , Vero 17 , Vero E6 18 , Huh-7 19 , and A549 14 cells, have been used for DENV infection. The antiviral agent ribavirin is currently used as the preferred treatment against infection by DENV 20 , 21 , ZIKV 21 , 22 , and hepatitis C virus (HCV) 23 .…”

Section: Discussionmentioning

confidence: 99%

Antiviral activity of pimecrolimus against dengue virus type 2 infection in vitro and in vivo

Kim,

Lee,

Kang

et al. 2024

Sci Rep

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Dengue virus (DENV) infection is a public health concern in several countries and is associated with severe diseases, such as dengue hemorrhagic fever and dengue shock syndrome. DENVs are transmitted to humans via the bites of infected Aedes mosquitoes, and no antiviral therapeutics are currently available. In this work, we aimed to identify antiviral drugs against DENV type 2 (DENV2) infections and selected pimecrolimus as a potential antiviral drug candidate. Pimecrolimus significantly inhibited DENV2-mediated cell death and replication in vitro. We also confirmed a decrease in the number of plaques formed as well as in the envelope protein levels of DENV2. The time-of-addition and course experiments revealed that pimecrolimus inhibited DENV2 infection during the early stages of the virus replication cycle. In an experimental mouse model, orally administered pimecrolimus alleviated body weight loss and lethality caused by DENV2 infection, which we used as readouts of the drug’s antiviral potency. Furthermore, pimecrolimus significantly inhibited the DENV2 load and ameliorated focal necrosis in the liver and spleen. Taken together, our in vitro and in vivo findings suggest that pimecrolimus is a promising antiviral drug candidate for the treatment of DENV2 infection.

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Section: Discussionmentioning

confidence: 99%

Antiviral activity of pimecrolimus against dengue virus type 2 infection in vitro and in vivo

Kim,

Lee,

Kang

et al. 2024

Sci Rep

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“…In addition, the cell suspensions collected in the kinetics were titrated by plaque assay on Vero cells using a protocol adapted from Malewicz and Jenkin, 1979 [ 32 ]. Titration was performed to compare the SLEV titer in the SLEV-positive control and co-infection conditions.…”

Section: Methodsmentioning

confidence: 99%

“…They were then observed daily under an inverted optical microscope, and when they showed a cytopathic effect, they were frozen at −80 • C in a freezer. After this, aliquots of 100 µL of this material were created in Eppendorf tubes to be stored in a freezer at −80 • C. Subsequently, the viral titer of SLEV was obtained by plaque assay, and the titer of BREV was obtained by TCID 50 [32,33]. To use the titer values (PFU/mL) in the MOI formula, we transformed the BREV titer from TCID 50 / mL to PFU/mL using the formula TCID 50 /mL titer × 0.7 = titer in PFU/mL [34].…”

Section: Preparation and Titration Of The Viral Stockmentioning

confidence: 99%

Viral Interference between the Insect-Specific Virus Brejeira and the Saint Louis Encephalitis Virus In Vitro

Ribeiro,

Martins,

Silva

et al. 2024

Viruses

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The Saint Louis encephalitis virus (SLEV) is an encephalitogenic arbovirus (Flaviviridae family) that has a wide geographical distribution in the western hemisphere, especially in the Americas. The negevirus Brejeira (BREV) was isolated for the first time in Brazil in 2005. This study aimed to verify the existence of a possible interfering effect of BREV on the course of SLEV infection and vice versa. We used clone C6/36 cells. Three combinations of MOIs were used (SLEV 0.1 × BREV 1; SLEV 1 × BREV 0.1; SLEV 1 × BREV 1) in the kinetics of up to 7 days and then the techniques of indirect immunofluorescence (IFA), a plaque assay on Vero cells, and RT-PCR were performed. Our results showed that the cytopathic effect (CPE) caused by BREV was more pronounced than the CPE caused by SLEV. Results of IFA, the plaque assay, and RT-PCR showed the suppression of SLEV replication in the co-infection condition in all the MOI combinations used. The SLEV suppression was dose-dependent. Therefore, the ISV Brejeira can suppress SLEV replication in Aedes albopictus cells, but SLEV does not negatively interfere with BREV replication.

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“…Two animals ( n = 2) were isolated to compose the control group (G1) and inoculated with a 0.5 mL suspension of strain-free VERO cells (ATCC number CCL-81, Washington, DC, USA). The other animals (G2; n = 3) were experimentally infected by the Asian genotype of ZIKV, strain BE H815744 (Genbank KU365780) isolated by Department of Arbovirology and Hemorrhagic Fevers, Evandro Chagas Institute, Ananindeua, Brazil, the inoculum with a 0.5 mL suspension of infected VERO cells containing 1.0 × 10 5 PFU/mL via intradermal injection at the level of the third right intercostal space below the right nipple [ 34 ] ( Figure 1 ).…”

Section: Methodsmentioning

confidence: 99%

Zika Virus Infection Damages the Testes in Pubertal Common Squirrel Monkeys (Saimiri collinsi)

Benchimol

Santos

Lopes³

et al. 2023

Viruses

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During the Zika virus (ZIKV) outbreak and after evidence of its sexual transmission was obtained, concerns arose about the impact of the adverse effects of ZIKV infection on human fertility. In this study, we evaluated the clinical-laboratory aspects and testicular histopathological patterns of pubertal squirrel monkeys (Saimiri collinsi) infected with ZIKV, analyzing the effects at different stages of infection. The susceptibility of S. collinsi to ZIKV infection was confirmed by laboratory tests, which detected viremia (mean 1.63 × 106 RNA copies/µL) and IgM antibody induction. Reduced fecal testosterone levels, severe testicular atrophy and prolonged orchitis were observed throughout the experiment by ultrasound. At 21 dpi, testicular damage associated with ZIKV was confirmed by histopathological and immunohistochemical (IHC) analyses. Tubular retraction, the degeneration and necrosis of somatic and germ cells in the seminiferous tubules, the proliferation of interstitial cells and an inflammatory infiltrate were observed. ZIKV antigen was identified in the same cells where tissue injuries were observed. In conclusion, squirrel monkeys were found to be susceptible to the Asian variant of ZIKV, and this model enabled the identification of multifocal lesions in the seminiferous tubules of the infected group evaluated. These findings may suggest an impact of ZIKV infection on male fertility.

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Development of dengue virus plaques under serum-free overlay medium

Cited by 19 publications

References 21 publications

Antiviral activity of pimecrolimus against dengue virus type 2 infection in vitro and in vivo

Antiviral activity of pimecrolimus against dengue virus type 2 infection in vitro and in vivo

Viral Interference between the Insect-Specific Virus Brejeira and the Saint Louis Encephalitis Virus In Vitro

Zika Virus Infection Damages the Testes in Pubertal Common Squirrel Monkeys (Saimiri collinsi)

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