An In Vitro Model of Latency and Reactivation of Varicella Zoster Virus in Human Stem Cell-Derived Neurons

Markus, Amos; Lebenthal-Loinger, Ilana; Yang, In Hong; Kinchington, Paul R.; Goldstein, Ronald S.

doi:10.1371/journal.ppat.1004885

Cited by 68 publications

(116 citation statements)

References 60 publications

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“…1D). The pattern of VZV gene expression was consistent between both cultures and shared many characteristics with quiescent VZV obtained from neuronal cultures using a different system (21).…”

Section: Vzv Infection Of the Soma Of Human Neurons Usually Induces Asupporting

confidence: 62%

“…Herpes zoster associated with varicella vaccine is most common at the site of the vaccine inoculation (43), and about 50% of zoster cases associated with vaccination are due to WT virus (50). Also, VZV has previously been shown to enter neurons in vitro from nerve endings by cell-to-cell contact with infected cells or cell-free virus followed by transaxonal transport to the soma (16,17,21).…”

Section: Discussionmentioning

confidence: 99%

“…Recently, Markus et al (21) reported an in vitro model of latency and reactivation of VZV in hESC-derived neurons using a compartmented microfluidic chamber and selective axonal infection of POka-based recombinant virus. A latent infection could be established with expression of multiple VZV mRNAs at low levels; induction of reactivation by a phosphoinositide 3-kinase (PI3K) inhibitor resulted in spreading of VZV ORF66 protein kinase-fused GFP within a cluster of neurons.…”

Section: Significancementioning

confidence: 99%

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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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Section: Vzv Infection Of the Soma Of Human Neurons Usually Induces Asupporting

confidence: 62%

Section: Discussionmentioning

confidence: 99%

Section: Significancementioning

confidence: 99%

See 1 more Smart Citation

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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“…In addition, human neurons derived from induced pluripotent stem cells (iPSCs) (29), neuronal stem cells (30), and embryonic stem cells (ESCs) (31,32) have been explored as infection models for human alphaherpesvirus. While these human cells support HSV productive infection (24,(27)(28)(29)31), reliable models to study the establishment of latency and/or reactivation have not been achieved using these human cells.…”

mentioning

confidence: 99%

“…In the presence of doxycycline, HD10.6 cells mature to exhibit a sensory neuron-associated phenotype on the basis of positive staining for neuronal cytoskeletal markers, expression of neuron-associated transcription factors, firing of action potentials, and capsaicin sensitivity (46). By infecting matured HD10.6 cells in the presence of acyclovir, we are able to describe a human cell culture model that exhibits experimental hallmarks of HSV-1 latency (15,17,18,32,47), including the presence of nonproductive viral genomes that retain the potential to reactivate and produce infectious virus. This model will enable the genetic and biochemical analysis of the molecular mechanisms of alphaherpesvirus latency and reactivation in human sensory neurons.…”

mentioning

confidence: 99%

An Immortalized Human Dorsal Root Ganglion Cell Line Provides a Novel Context To Study Herpes Simplex Virus 1 Latency and Reactivation

Thellman

Botting

Madaj

et al. 2017

J Virol

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A defining characteristic of alphaherpesviruses is the establishment of lifelong latency in host sensory ganglia with occasional reactivation causing recurrent lytic infections. As an alternative to rodent models, we explored the use of an immortalized cell line derived from human dorsal root ganglia. HD10.6 cells proliferate by virtue of a transduced tetracycline-regulated v-myc oncogene. In the presence of doxycycline, HD10.6 cells mature to exhibit neuronal morphology and express sensory neuron-associated markers such as neurotrophin receptors TrkA, TrkB, TrkC, and RET and the sensory neurofilament peripherin. Infection of mature HD10.6 neurons by herpes simplex virus 1 (HSV-1) results in a delayed but productive infection. However, infection at a low multiplicity of infection (MOI) in the presence of acyclovir results in a quiescent infection resembling latency in which viral genomes are retained in a low number of neurons, viral gene expression is minimal, and infectious virus is not released. At least some of the quiescent viral genomes retain the capacity to reactivate, resulting in viral DNA replication and release of infectious virus. Reactivation can be induced by depletion of nerve growth factor; other commonly used reactivation stimuli have no significant effect.IMPORTANCE Infections by herpes simplex viruses (HSV) cause painful cold sores or genital lesions in many people; less often, they affect the eye or even the brain. After the initial infection, the virus remains inactive or latent in nerve cells that sense the region where that infection occurred. To learn how virus maintains and reactivates from latency, studies are done in neurons taken from rodents or in whole animals to preserve the full context of infection. However, some cellular mechanisms involved in HSV infection in rodents are different from those in humans. We describe the use of a human cell line that has the properties of a sensory neuron. HSV infection in these cultured cells shows the properties expected for a latent infection, including reactivation to produce newly infectious virus. Thus, we now have a cell culture model for latency that is derived from the normal host for this virus.KEYWORDS HSV-1, quiescent infection, sensory neurons H erpesviruses are unusual among viruses in that they employ two infection strategies, lytic (replicative or productive) infection and latency (quiescent) infection. Alphaherpesviruses typically replicate in epithelial cells and establish latency in the peripheral nervous system within the sensory neurons of the ganglia that innervate mucosa of the primary infection site (1), such as the trigeminal ganglia (TG) and dorsal root ganglia (DRG). Operationally, latency is defined as "the persistence of a viral genome within tissue where, at any given time, there is a population of cells that lack detectable infectious virus, viral proteins, or viral lytic transcripts that are dormant but have the capability of being reactivated" (2). During latency of herpes simplex viruses

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Organoids: development and applications in disease models, drug discovery, precision medicine, and regenerative medicine

Yao,

Cheng,

Pan

et al. 2024

MedComm

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Organoids are miniature, highly accurate representations of organs that capture the structure and unique functions of specific organs. Although the field of organoids has experienced exponential growth, driven by advances in artificial intelligence, gene editing, and bioinstrumentation, a comprehensive and accurate overview of organoid applications remains necessary. This review offers a detailed exploration of the historical origins and characteristics of various organoid types, their applications—including disease modeling, drug toxicity and efficacy assessments, precision medicine, and regenerative medicine—as well as the current challenges and future directions of organoid research. Organoids have proven instrumental in elucidating genetic cell fate in hereditary diseases, infectious diseases, metabolic disorders, and malignancies, as well as in the study of processes such as embryonic development, molecular mechanisms, and host–microbe interactions. Furthermore, the integration of organoid technology with artificial intelligence and microfluidics has significantly advanced large‐scale, rapid, and cost‐effective drug toxicity and efficacy assessments, thereby propelling progress in precision medicine. Finally, with the advent of high‐performance materials, three‐dimensional printing technology, and gene editing, organoids are also gaining prominence in the field of regenerative medicine. Our insights and predictions aim to provide valuable guidance to current researchers and to support the continued advancement of this rapidly developing field.

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An In Vitro Model of Latency and Reactivation of Varicella Zoster Virus in Human Stem Cell-Derived Neurons

Cited by 68 publications

References 60 publications

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

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

An Immortalized Human Dorsal Root Ganglion Cell Line Provides a Novel Context To Study Herpes Simplex Virus 1 Latency and Reactivation

Organoids: development and applications in disease models, drug discovery, precision medicine, and regenerative medicine

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