Application of quantitative immunofluorescence assays to analyze the expression of cell contact proteins during Zika virus infections

Leiva, Santiago; Dizanzo, María Paula; Fabbri, Cintia; Valdano, Marina Bugnon; Luppo, Victoria; Levis, Silvana; Cavatorta, Ana Laura; Morales, María Alejandra; Gardiol, Daniela

doi:10.1016/j.virusres.2021.198544

Cited by 2 publications

(1 citation statement)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Then, each slide was incubated with diluted primary antibody ICP5 (Abcam, Cambridge, UK; 1:500) in a humidified chamber box overnight at 4 °C. Subsequently, the slides were washed with PBST and incubated with FITC-labeled fluorescent secondary antibody (Proteintech, Chicago, CA, USA; 1:200) at 37 °C for 1 h. Finally, the slides were sealed by dropping the sealing liquid containing DAPI and an anti-fluorescence quencher, and the images were observed and collected using a panoramic MIDI digital scanner (3D HISTECH, Budapest, Hungary), as described previously [ 53 ].…”

Section: Methodsmentioning

confidence: 99%

Metabolomics Profiles Reveal New Insights of Herpes Simplex Virus Type 1 Infection

Huang

Wang

Lei

et al. 2023

IJMS

View full text Add to dashboard Cite

Herpes simplex virus type 1 (HSV-1) is a ubiquitous human pathogen that can cause significant morbidity, primarily facial cold sores and herpes simplex encephalitis. Previous studies have shown that a variety of viruses can reprogram the metabolic profiles of host cells to facilitate self-replication. In order to further elucidate the metabolic interactions between the host cell and HSV-1, we used liquid chromatography-tandem mass spectrometry (LC-MS/MS) to analyze the metabolic profiles in human lung fibroblasts KMB17 infected with HSV-1. The results showed that 654 and 474 differential metabolites were identified in positive and negative ion modes, respectively, and 169 and 114 metabolic pathways that might be altered were screened. These altered metabolites are mainly involved in central carbon metabolism, choline metabolism, amino acid metabolism, purine and pyrimidine metabolism, cholesterol metabolism, bile secretion, and prolactin signaling pathway. Further, we confirmed that the addition of tryptophan metabolite kynurenine promotes HSV-1 replication, and the addition of 25-Hydroxycholesterol inhibits viral replication. Significantly, HSV-1 replication was obviously enhanced in the ChOKα (a choline metabolic rate-limiting enzyme) deficient mouse macrophages. These results indicated that HSV-1 induces the metabolic reprogramming of host cells to promote or resist viral replication. Taken together, these observations highlighted the significance of host cell metabolism in HSV-1 replication, which would help to clarify the pathogenesis of HSV-1 and identify new anti-HSV-1 therapeutic targets.

show abstract

Section: Methodsmentioning

confidence: 99%

Metabolomics Profiles Reveal New Insights of Herpes Simplex Virus Type 1 Infection

Huang

Wang

Lei

et al. 2023

IJMS

View full text Add to dashboard Cite

show abstract

Dapoxetine, a Selective Serotonin Reuptake Inhibitor, Suppresses Zika Virus Infection In Vitro

Zhang,

Yu,

Zhu

et al. 2023

Molecules

View full text Add to dashboard Cite

Zika virus (ZIKV) belongs to the Flavivirus genus of the Flaviviridae family, and is a pathogen posing a significant threat to human health. Currently, there is a lack of internationally approved antiviral drugs for the treatment of ZIKV infection, and symptomatic management remains the primary clinical approach. Consequently, the exploration of safe and effective anti-ZIKV drugs has emerged as a paramount imperative in ZIKV control efforts. In this study, we performed a screening of a compound library consisting of 1789 FDA-approved drugs to identify potential agents with anti-ZIKV activity. We have identified dapoxetine, an orally administered selective serotonin reuptake inhibitor (SSRI) commonly employed for the clinical management of premature ejaculation (PE), as a potential inhibitor of ZIKV RNA-dependent RNA polymerase (RdRp). Consequently, we conducted surface plasmon resonance (SPR) analysis to validate the specific binding of dapoxetine to ZIKV RdRp, and further evaluated its inhibitory effect on ZIKV RdRp synthesis using the ZIKV Gluc reporter gene assay. Furthermore, we substantiated the efficacy of dapoxetine in suppressing intracellular replication of ZIKV, thereby demonstrating a concentration-dependent antiviral effect (EC50 values ranging from 4.20 μM to 12.6 μM) and negligible cytotoxicity (CC50 > 50 μM) across diverse cell lines. Moreover, cell fluorescence staining and Western blotting assays revealed that dapoxetine effectively reduced the expression of ZIKV proteins. Collectively, our findings suggest that dapoxetine exhibits anti-ZIKV effects by inhibiting ZIKV RdRp activity, positioning it as a potential candidate for clinical therapeutic intervention against ZIKV infection.

show abstract

Application of quantitative immunofluorescence assays to analyze the expression of cell contact proteins during Zika virus infections

Cited by 2 publications

References 42 publications

Metabolomics Profiles Reveal New Insights of Herpes Simplex Virus Type 1 Infection

Metabolomics Profiles Reveal New Insights of Herpes Simplex Virus Type 1 Infection

Dapoxetine, a Selective Serotonin Reuptake Inhibitor, Suppresses Zika Virus Infection In Vitro

Contact Info

Product

Resources

About