Electric Cell-Substrate Impedance Sensing To Monitor Viral Growth and Study Cellular Responses to Infection with Alphaherpesviruses in Real Time

Pennington, Matthew R.; Walle, Gerlinde R. Van de

doi:10.1128/msphere.00039-17

Cited by 31 publications

(30 citation statements)

References 41 publications

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“…Influenza A virus genome copies varied among different sample types with fomites having the highest copy number followed by air 1–4 μm particles ( S1 Table ). The electrical impedance assay has been used by other groups [ 21 , 22 ] for assessing influenza A infectivity and for other viral infections [ 23 , 24 ]. The instrument measures cell index as a read out of cell adherence, viability, and growth.…”

Section: Resultsmentioning

confidence: 99%

Humidity as a non-pharmaceutical intervention for influenza A

et al. 2018

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Influenza is a global problem infecting 5–10% of adults and 20–30% of children annually. Non-pharmaceutical interventions (NPIs) are attractive approaches to complement vaccination in the prevention and reduction of influenza. Strong cyclical reduction of absolute humidity has been associated with influenza outbreaks in temperate climates. This study tested the hypothesis that raising absolute humidity above seasonal lows would impact influenza virus survival and transmission in a key source of influenza virus distribution, a community school. Air samples and objects handled by students (e.g. blocks and markers) were collected from preschool classrooms. All samples were processed and PCR used to determine the presence of influenza virus and its amount. Additionally samples were tested for their ability to infect cells in cultures. We observed a significant reduction (p < 0.05) in the total number of influenza A virus positive samples (air and fomite) and viral genome copies upon humidification as compared to control rooms. This suggests the future potential of artificial humidification as a possible strategy to control influenza outbreaks in temperate climates. There were 2.3 times as many ILI cases in the control rooms compared to the humidified rooms, and whether there is a causal relationship, and its direction between the number of cases and levels of influenza virus in the rooms is not known. Additional research is required, but this is the first prospective study suggesting that exogenous humidification could serve as a scalable NPI for influenza or other viral outbreaks.

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Section: Resultsmentioning

confidence: 99%

Humidity as a non-pharmaceutical intervention for influenza A

et al. 2018

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“…This leads to the formation of herpes stromal keratitis (HSK), which consists of severe corneal lesions, corneal neovascularization and infiltration of immune cells that contribute to destruction of the cornea, ultimately leading to blindness. Due to the striking similarities in pathogenesis of HSV-1, CHV-1 and FHV-1, studies in dogs and cats are proposed as valuable virus-natural host models to study the pathogenesis of human herpesvirus infections of the eye [ 2 , 5 , 8 , 9 ].…”

Section: Introductionmentioning

confidence: 99%

New Paradigms for the Study of Ocular Alphaherpesvirus Infections: Insights into the Use of Non-Traditional Host Model Systems

Pennington

Ledbetter

Walle

2017

Viruses

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Ocular herpesviruses, most notably human alphaherpesvirus 1 (HSV-1), canid alphaherpesvirus 1 (CHV-1) and felid alphaherpesvirus 1 (FHV-1), infect and cause severe disease that may lead to blindness. CHV-1 and FHV-1 have a pathogenesis and induce clinical disease in their hosts that is similar to HSV-1 ocular infections in humans, suggesting that infection of dogs and cats with CHV-1 and FHV-1, respectively, can be used as a comparative natural host model of herpesvirus-induced ocular disease. In this review, we discuss both strengths and limitations of the various available model systems to study ocular herpesvirus infection, with a focus on the use of these non-traditional virus-natural host models. Recent work has demonstrated the robustness and reproducibility of experimental ocular herpesvirus infections in dogs and cats, and, therefore, these non-traditional models can provide additional insights into the pathogenesis of ocular herpesvirus infections.

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“…Next, we wanted to confirm this downregulation of late gene expression in raltegravir-treated FeHV-1-infected cells on the protein level. However, since monoclonal antibodies targeting single FeHV-1 proteins are not commercially available, we utilized a recombinant FeHV-1 strain expressing the DsRed Express2 fluorophore fused to the C terminus of glycoprotein D (FeHV-1-gD-DsRed) (48). The virus was suitable for these experiments, as gD expression was found to be downregulated by raltegravir, but not PAA, treatment ( Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The FeHV-1 strain FH2CS (63) was used for all experiments and compared with the FeHV-1 strain C-27 (64) for sequencing and drug susceptibility purposes. For flow cytometry, a recombinant FH2CS FeHV-1 strain (FeHV-1-gD-DsRed) was used, which we generated in house using clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 genome engineering to express DsRed Express 2 fused to the C-terminal end of glycoprotein D (48). All viral stocks were grown and their titers were determined on CRFK cells, as previously described (26).…”

Section: Cells Viruses and Drugs Crfk Cells (American Typementioning

confidence: 99%

The HIV Integrase Inhibitor Raltegravir Inhibits Felid Alphaherpesvirus 1 Replication by Targeting both DNA Replication and Late Gene Expression

Pennington

Voorhees

Callaway

et al. 2018

J Virol

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Alphaherpesvirus-associated ocular infections in humans, caused by human alphaherpesvirus 1 (HHV-1), remain challenging to treat due to the frequency of drug application required and the potential for the selection of drug resistant viruses. Repurposing on-the-market drugs is a viable strategy to accelerate the pace of drug development. It has been reported that the human immunodeficiency virus (HIV) integrase inhibitor raltegravir inhibits HHV-1 replication by targeting the DNA polymerase accessory factor and limits terminase-mediated genome cleavage of the human betaherpesvirus 5 (HHV-5). We have previously shown, both and that raltegravir can also inhibit the replication of felid alphaherpesvirus 1 (FeHV-1), a common ocular pathogen of cats with a similar pathogenesis to HHV-1 ocular disease. In contrast to what was reported for HHV-1, we were unable to select for a raltegravir-resistant FeHV-1 in order to define any basis for drug action. A candidate-based approach to explore the mode-of-action of raltegravir against FeHV-1 showed that raltegravir did not impact FeHV-1 terminase function, as described for HHV-5. Instead, raltegravir inhibited DNA replication, similar to HHV-1, but by targeting the initiation of viral DNA replication rather than elongation. In addition, we found that raltegravir specifically repressed late gene expression independent of DNA replication, and both activities are consistent with inhibition of ICP8. Taken together, these results suggest that raltegravir could be a valuable therapeutic agent against herpesviruses. The rise of drug-resistant herpesviruses is a long-standing concern, particularly among immunocompromised patients. Therefore, therapies targeting viral proteins other than the DNA polymerase that may be less likely to lead to drug-resistant viruses are urgently needed. Using FeHV-1, an alphaherpesvirus closely related to HHV-1 that similarly causes ocular herpes in its natural host, we found that the HIV integrase inhibitor raltegravir targets different stages of the virus life cycle beyond DNA replication and that it does so without developing drug resistance under the conditions tested. This shows that this drug could prove a viable strategy for the treatment of herpesvirus infections.

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Electric Cell-Substrate Impedance Sensing To Monitor Viral Growth and Study Cellular Responses to Infection with Alphaherpesviruses in Real Time

Cited by 31 publications

References 41 publications

Humidity as a non-pharmaceutical intervention for influenza A

Humidity as a non-pharmaceutical intervention for influenza A

New Paradigms for the Study of Ocular Alphaherpesvirus Infections: Insights into the Use of Non-Traditional Host Model Systems

The HIV Integrase Inhibitor Raltegravir Inhibits Felid Alphaherpesvirus 1 Replication by Targeting both DNA Replication and Late Gene Expression

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