Molecular Characterization of Varicella Zoster Virus in Latently Infected Human Ganglia: Physical State and Abundance of VZV DNA, Quantitation of Viral Transcripts and Detection of VZV-Specific Proteins

Azarkh, Yevgeniy; Gilden, Don; Cohrs, Randall J.

doi:10.1007/82_2009_2

Cited by 21 publications

(22 citation statements)

References 44 publications

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“…In contrast, VZV gene expression during productive replication in a fully permissive cell type follows an ordered cascade, with the expression of immediate-early, early, and late genes in a sequential manner (28,35). VZV glycoproteins (late gene products) are not expressed during latent VZV infection (6), and thus, their detection in neurons provides a useful means to exclude true latency. We therefore examined ganglionic sections from cases following herpes zoster for VZV gE by IHC staining.…”

Section: Resultsmentioning

confidence: 99%

Characterization of the Host Immune Response in Human Ganglia after Herpes Zoster

Gowrishankar

Steain

Cunningham

et al. 2010

J Virol

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Varicella-zoster virus (VZV) causes varicella (chicken pox) and establishes latency in ganglia, from where it reactivates to cause herpes zoster (shingles), which is often followed by postherpetic neuralgia (PHN), causing severe neuropathic pain that can last for years after the rash. Despite the major impact of herpes zoster and PHN on quality of life, the nature and kinetics of the virus-immune cell interactions that result in ganglion damage have not been defined. We obtained rare material consisting of seven sensory ganglia from three donors who had suffered from herpes zoster between 1 and 4.5 months before death but who had not died from herpes zoster. We performed immunostaining to investigate the site of VZV infection and to phenotype immune cells in these ganglia. VZV antigen was localized almost exclusively to neurons, and in at least one case it persisted long after resolution of the rash. The large immune infiltrate consisted of noncytolytic CD8؉ T cells, with lesser numbers of CD4 ؉ T cells, B cells, NK cells, and macrophages and no dendritic cells. VZV antigen-positive neurons did not express detectable major histocompatibility complex (MHC) class I, nor did CD8 ؉ T cells surround infected neurons, suggesting that mechanisms of immune control may not be dependent on direct contact. This is the first report defining the nature of the immune response in ganglia following herpes zoster and provides evidence for persistence of non-latency-associated viral antigen and inflammation beyond rash resolution.Varicella-zoster virus (VZV) is a highly species-specific human alphaherpesvirus that infects a majority of the world's population. VZV causes two clinically significant diseases; varicella (chicken pox) and herpes zoster (shingles) (5,8,19). Varicella is characterized by widespread cutaneous vesicular lesions and is a consequence of primary VZV infection in VZV-naïve individuals. While varicella is a relatively mild disease in immunocompetent children, it can cause significant morbidity in healthy adults and is frequently life threatening in immunocompromised individuals (3,4,22). The innate and adaptive immune responses act to eliminate replicating virus during varicella, but not all virus is cleared during this time, with some presumed to access nerve axons in the skin, enabling transport to neurons in sensory ganglia, where the virus is able to establish a lifelong latent infection (5,8,12,13,20,32). An alternative possibility is that virus is transported to ganglia via hematogenous spread (36). The ability of VZV to establish latency in the host is critical to the success of this virus as a human pathogen.VZV reactivation from latency (herpes zoster) causes serious disease in older and immunocompromised individuals and is characterized by vesicular skin rash in a dermatomal distribution with preceding and concomitant pain (7,10,21,68). During reactivation, sensory ganglia are sites of viral replication, where an intense inflammatory response is induced and widespread necrosis of glial cells and neur...

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Section: Resultsmentioning

confidence: 99%

Characterization of the Host Immune Response in Human Ganglia after Herpes Zoster

Gowrishankar

Steain

Cunningham

et al. 2010

J Virol

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“…Both VZV 63 and VZV gE proteins were found exclusively in neurons, but not in nonneuronal cells. Importantly, transcripts corresponding to VZV ORFs 21, 29, 62, 63, and 68 (gE) are present in latently infected human ganglia (1,19). VZV gE is also present in vivo months after experimental infection of human dorsal root ganglia (22).…”

Section: Discussionmentioning

confidence: 99%

Varicella-Zoster Virus Infection of Differentiated Human Neural Stem Cells

Pugazhenthi

Nair

Velmurugan

et al. 2011

J Virol

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Primary varicella-zoster virus (VZV) infection in humans produces varicella (chickenpox),Varicella zoster virus (VZV) is a ubiquitous, exclusively human, and highly neurotropic herpesvirus. Primary infection causes varicella (chickenpox), during which VZV replicates in the cells of every visceral organ and the skin (12). VZV establishes lifelong latency in neurons of cranial nerve ganglia, dorsal root ganglia, and autonomic ganglia along the entire neuraxis (9,11,17). VZV reactivates in elderly and immunocompromised individuals to produce shingles (herpes zoster), frequently complicated by chronic pain (postherpetic neuralgia) (10) and other serious neurological and ocular disorders, such as VZV vasculopathy, myelopathy, and retinal necrosis.Analyses of latently infected human ganglia obtained postmortem within 24 h after death have provided useful information on the configuration and abundance of the VZV genome and the limited extent of viral gene expression (reviewed in reference 6). However, such studies are hampered by lack of sufficient tissue, RNA degradation postmortem, and possible virus reactivation after death. Because no satisfactory animal model exists to study the interaction of VZV with neurons, alternate models have been developed. For example, immunedeficient SCID mice have had human neural stem cells or human ganglia transplanted, followed by infection with VZV and examination of infected neurons (2, 21). VZV has been also shown to infect cultured neurons from guinea pig enteric ganglia (8). An in vitro model of cultured human neurons infected with VZV provides an opportunity to study the molecular events leading to the establishment of VZV latency and reactivation. Here, we developed a system in which human neural stem cells (NSCs) were induced to differentiate into neurons, infected with VZV, observed carefully, and analyzed for viral gene expression and for markers of apoptosis.

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“…Human cadaveric ganglia have been used to study viral transcripts and proteins expressed during latency. Although several VZV mRNAs and proteins have been detected in human ganglia (12), more recent studies suggest that viral protein expression during latency observed in certain prior studies may be due to nonspecific Ab staining (13,14), and viral mRNA expression has been postulated to reflect postmortem events rather than the true latent state (15). Recent advances in stem cell research allow a virtually unlimited supply of human neurons differentiated from hESCs or human-induced pluripotent stem cells that have not been infected with VZV.…”

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confidence: 99%

In vitro system using human neurons demonstrates that varicella-zoster vaccine virus is impaired for reactivation, but not latency

Sadaoka

Depledge

Rajbhandari

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

104

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Varicella-zoster virus (VZV) establishes latency in human sensory and cranial nerve ganglia during primary infection (varicella), and the virus can reactivate and cause zoster after primary infection. The mechanism of how the virus establishes and maintains latency and how it reactivates is poorly understood, largely due to the lack of robust models. We found that axonal infection of neurons derived from hESCs in a microfluidic device with cell-free parental Oka (POka) VZV resulted in latent infection with inability to detect several viral mRNAs by reverse transcriptase-quantitative PCR, no production of infectious virus, and maintenance of the viral DNA genome in endless configuration, consistent with an episome configuration. With deep sequencing, however, multiple viral mRNAs were detected. Treatment of the latently infected neurons with Ab to NGF resulted in production of infectious virus in about 25% of the latently infected cultures. Axonal infection of neurons with vaccine Oka (VOka) VZV resulted in a latent infection similar to infection with POka; however, in contrast to POka, VOka-infected neurons were markedly impaired for reactivation after treatment with Ab to NGF. In addition, viral transcription was markedly reduced in neurons latently infected with VOka compared with POka. Our in vitro system recapitulates both VZV latency and reactivation in vivo and may be used to study viral vaccines for their ability to establish latency and reactivate. varicella-zoster virus | varicella vaccine | reactivation | latency | herpesvirus V aricella-zoster virus (VZV) is a member of Alphaherpesvirinae, characterized by neurotropism and lifelong latent infection in human dorsal root, cranial nerve, and enteric ganglia (1). During primary infection (varicella), VZV gains access to sensory neurons by two potential routes: retrograde axonal transport from cutaneous lesions or infection of neurons by virus-infected T cells circulating through the body (2). The virus then establishes latency in neurons, and months to years later, when VZV-specific T cells decline, the virus can reactivate to cause herpes zoster.VZV is the only human herpesvirus for which a licensed vaccine is approved, and the live-attenuated vaccine Oka (VOka) virus is effective in preventing both varicella and zoster. VOka was derived from WT parental Oka (POka) by propagation in human embryonic lung cells, guinea pig embryo fibroblasts, and human fibroblasts (3, 4). VOka is a genetic mixture of at least eight genotypes (5), and, initially, the genomes of POka and VOka were found to differ by 42 nucleotides and 20 amino acids (6). A recent study using deep sequencing identified 165 additional nucleotide changes in VOka, although most changes were presence in <10% of viral genomes (7). Attenuation of VOka is postulated to be due to mutations in multiple regions of the genome (8, 9).Despite numerous studies, the mechanisms for establishment and maintenance of VZV latency and subsequent reactivation remain poorly understood, primarily due to the lack of ...

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Molecular Characterization of Varicella Zoster Virus in Latently Infected Human Ganglia: Physical State and Abundance of VZV DNA, Quantitation of Viral Transcripts and Detection of VZV-Specific Proteins

Cited by 21 publications

References 44 publications

Characterization of the Host Immune Response in Human Ganglia after Herpes Zoster

Characterization of the Host Immune Response in Human Ganglia after Herpes Zoster

Varicella-Zoster Virus Infection of Differentiated Human Neural Stem Cells

In vitro system using human neurons demonstrates that varicella-zoster vaccine virus is impaired for reactivation, but not latency

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