A VP26-mNeonGreen Capsid Fusion HSV-2 Mutant Reactivates from Viral Latency in the Guinea Pig Genital Model with Normal Kinetics

Pieknik, Julianna R; Bertke, Andrea S.; Tang, Shuang; Krause, Philip R.

doi:10.3390/v10050246

Cited by 4 publications

(8 citation statements)

References 25 publications

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“…We used three different approaches to investigate the ability of HSV-2 to reactivate from autonomic ganglia, and we consistently found that the virus is latent and reactivation competent in both DRG and SSG. Given previous experiments that revealed LAT and viral DNA in the autonomic ganglia, viral reactivation in ganglia after an axotomy-induced neural stress signal is not unexpected (4,15,22). However, this is the first description of HSV-2 reactivation from latently infected autonomic ganglia in a clinically relevant vaginal model of infection, in which virus reactivates spontaneously to cause recurrent disease.…”

Section: Discussionmentioning

confidence: 63%

“…Autonomic and sensory ganglia harbor reactivation-competent HSV-2. To detect reactivation-competent latent virus in the dorsal root ganglia (DRG) and the sacral sympathetic ganglion (SSG), we infected guinea pigs intravaginally with "Nedel," a fluorescent mNeonGreen minor capsid fusion (VP26) HSV-2 recombinant virus that expresses the VP26-mNeonGreen fusion protein only during productive infection (22). Nedel replicates with wild-type kinetics and reactivates from latency in vivo.…”

Section: Resultsmentioning

confidence: 99%

“…HSV-2 strain 333 was originally obtained from Gary Hayward (Johns Hopkins University, Baltimore, MD). mNeonGreen was obtained from Allele Biotech (San Diego, CA), and the recombinant fluorescent virus Nedel has recently been described (22). Nedel expresses mNeonGreen as a fusion protein with VP26 during productive infection.…”

Section: Methodsmentioning

confidence: 99%

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Herpes Simplex Virus 2 in Autonomic Ganglia: Evidence for Spontaneous Reactivation

Pieknik

Bertke

Krause

2019

J Virol

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Herpes simplex virus 2 (HSV-2) can be transmitted in the presence or absence of lesions, allowing efficient spread among the general population. Recurrent HSV genital lesions are thought to arise from reactivated latent virus in sensory cell bodies of the dorsal root ganglia (DRG). However, HSV-2 has also been found latent in autonomic ganglia. Spontaneous reactivation or a low level of chronic infection could theoretically also occur in these peripheral nervous tissues, contributing to the presence of infectious virus in the periphery and to viral transmission. Use of a recently described, optimized virus with a monomeric mNeonGreen protein fused to viral capsid protein 26 (VP26) permitted detection of reactivating virus in explanted ganglia and cryosections of DRG and the sacral sympathetic ganglia (SSG) from latently infected guinea pigs. Immediate early, early, and late gene expression were quantified by droplet digital reverse transcription-PCR (ddRT-PCR), providing further evidence of viral reactivation not only in the expected DRG but also in the sympathetic SSG. These findings indicate that viral reactivation from autonomic ganglia is a feature of latent viral infection and that these reactivations likely contribute to viral pathogenesis. IMPORTANCE HSV-2 is a ubiquitous important human pathogen that causes recurrent infections for the life of its host. We hypothesized that the autonomic ganglia have important roles in viral reactivation, and this study sought to determine whether this is correct in the clinically relevant guinea pig vaginal infection model. Our findings indicate that sympathetic ganglia are sources of reactivating virus, helping explain how the virus causes lifelong recurrent disease.

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Section: Discussionmentioning

confidence: 63%

Section: Resultsmentioning

confidence: 99%

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Herpes Simplex Virus 2 in Autonomic Ganglia: Evidence for Spontaneous Reactivation

Pieknik

Bertke

Krause

2019

J Virol

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“…HSV-2 (ATCC VR-1781) (ATCC, Manassas, VA, USA) and HSV-2 VP26-GFP [ 40 ] which expresses a fusion protein of VP26 and GFP (generously donated by Dr. Andrea Bertke, Virginia Tech University) were used in all experiments. Preparation of virus was previously described in De Oliveira et al [ 37 ].…”

Section: Methodsmentioning

confidence: 99%

In Vitro and In Silico Analysis of the Inhibitory Activity of EGCG-Stearate against Herpes Simplex Virus-2

et al. 2022

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About half a billion people worldwide are infected with herpes simplex virus-2 (HSV-2). Prolonged treatment with acyclovir (ACV) and its analogs leads to the development of resistant strains. The aim of this study was to investigate the antiviral potential of epigallocatechin gallate (EGCG) from Camellia sinensis and a stable analog EGCG-stearate (EGCG-S) against HSV-2 in cultured Vero cells. Cell viability and cell proliferation assays were used to determine the non-cytotoxic concentrations on cultured Vero cells. HSV-2 with a green fluorescent protein (GFP) fusion protein of VP26 virions were treated with non-cytotoxic concentrations of EGCG and EGCG-S. The effects on infectivity and mechanisms were determined by plaque assay, attachment and penetration assays, confocal microscopy, qPCR, and in silico modeling analysis. Our results demonstrate that treatment of HSV-2 virions with EGCG and EGCG-S at a concentration of 75 µM showed greater than 99.9% inhibition by inhibiting the attachment of HSV-2 virions to host cells. The bioinformatic analysis indicated high binding affinity of EGCG-S for glycoprotein D; thus EGCG-S may block fusion of HSV-2 and the cell membrane, preventing entry of HSV-2 into the cell.

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“…Additionally, the idea that the reactivating neuron must survive based on this level of shedding does not appear to consider the number of ganglia that could be potentially alternate sources for the viral DNA in genital skin or mucosa. Progress in developing animal models to gain insight into the contribution of other ganglia to vaginal reactivation (Bertke et al, 2007;Lee et al, 2015;Pieknik et al, 2019;Yanez et al, 2017) as well as developing the tools in HSV2 to test hypotheses in vivo (Kawamura et al, 2018;Pieknik et al, 2018) will undoubtedly deepen current understanding of HSV1 and 2 reactivation and the distinctions between them.…”

Section: Frequency Of Hsv2 Genital Shedding Implies Hsv2 Is Continuously "Reactivating" In the Ganglion Corollary: The Reactivating Neuromentioning

confidence: 99%

Alphaherpesvirus Latency and Reactivation with a Focus on Herpes Simplex Virus

Sawtell¹,

Thompson²

2021

Current Issues in Molecular Biology

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We are at an interesting time in the understanding of alpha herpesvirus latency and reactivation and their implications to human disease. Conceptual advances have come from both animal and neuronal culture models. This review focuses on the concept that the tegument protein and viral transactivator VP16 plays a major role in the transition from latency to the lytic cycle. During acute infection, regulation of VP16 transactivation balances spread in the nervous system, establishment of latent infections and virulence. Reactivation is dependent on this transactivator to drive entry into the lytic cycle. In vivo de novo expression of VP16 protein is mediated by sequences conferring pre-immediate early transcription embedded in the normally leaky late promoter. In vitro, alternate mechanisms regulating VP16 expression in the context of latency have come from the SCG neuron culture model and include the concepts that (i) generalized transcriptional derepression of the caister.com/cimb 267 Curr. Issues Mol. Biol. Vol. 41 Latency and Reactivation Sawtell and Thompson viral genome and sequestration of VP16 in the cytoplasm for ~48 hours (Phase I) precedes and is required for VP16-dependent reactivation (Phase II); and (ii) a histone methyl/phospho switch during Phase I is required for Phase II reactivation. The challenge to the field is reconciling these data into a unified model of virus reactivation.The greatest enemy of knowledge is not ignorance, it is the illusion of knowledge.--Daniel J. Boorstin.The task of compiling this review was uncomfortably humbling, as if cataloging the stars in the universe. While not completely dark, our night sky is missing a multitude of studies which are among the many points of light contributing to our field. This article is a focused review in which we discuss from the vantage point of our expertise, just a handful of concepts that have or are emerging. A lookback at some of the pioneering work that grounds our field is also included.

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A VP26-mNeonGreen Capsid Fusion HSV-2 Mutant Reactivates from Viral Latency in the Guinea Pig Genital Model with Normal Kinetics

Cited by 4 publications

References 25 publications

Herpes Simplex Virus 2 in Autonomic Ganglia: Evidence for Spontaneous Reactivation

Herpes Simplex Virus 2 in Autonomic Ganglia: Evidence for Spontaneous Reactivation

In Vitro and In Silico Analysis of the Inhibitory Activity of EGCG-Stearate against Herpes Simplex Virus-2

Alphaherpesvirus Latency and Reactivation with a Focus on Herpes Simplex Virus

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