Varicella-Zoster Virus Infects Human Embryonic Stem Cell-Derived Neurons and Neurospheres but Not Pluripotent Embryonic Stem Cells or Early Progenitors

Dukhovny, Anna; Sloutskin, Anna; Markus, Amos; Yee, Michael B.; Kinchington, Paul R.; Goldstein, Ronald S.

doi:10.1128/jvi.06810-11

Cited by 35 publications

(39 citation statements)

References 28 publications

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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%

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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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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“…In contrast, hESC were susceptible to herpes simplex virus 1 (HSV-1) and adenovirus (data not shown). These findings, along with reports of their susceptibility to HSV-1, pseudorabies virus (PrV), coxsackie virus, and varicella-zoster virus (VZV), when grown in suspension (34,35), suggest a virus-specific restriction mechanism(s).…”

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confidence: 99%

Transition toward Human Cytomegalovirus Susceptibility in Early Human Embryonic Stem Cell-Derived Neural Precursors

Berger

Gil

Panet

et al. 2015

J Virol

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Congenital human cytomegalovirus (HCMV) infection is associated with neurodevelopmental disabilities. To dissect the earliest events of infection in the developing human brain, we studied HCMV infection during controlled differentiation of human embryonic stem cells (hESC) into neural precursors. We traced a transition from viral restriction in hESC, mediated by a block in viral binding, toward HCMV susceptibility in early hESC-derived neural precursors. We further revealed the role of plateletderived growth factor receptor alpha (PDGFR␣) as a determinant of the developmentally acquired HCMV susceptibility. Human cytomegalovirus (HCMV) is a leading cause of congenital infection (1), associated with neurodevelopmental disabilities (2, 3). The ability of the virus to infect the developing fetal brain is a key factor in its neuropathogenesis (3)(4)(5)(6)(7)(8). While considerable experimental data were obtained from newborn and embryonic mouse models (6, 9-15), the strict species specificity precludes animal models of HCMV. Recent studies in human neuronal progenitor cells (NPC) derived from fetal and neonatal brains have revealed productive HCMV infection of NPC, with resultant functional alterations (16)(17)(18)(19)(20). Notably, NPC do not represent early neural embryonic development but rather a more advanced stage along the neuronal differentiation route. Hence, the earliest events defining the susceptibility of the developing human nervous system to HCMV have remained largely unknown.Human embryonic stem cells (hESC) provide an opportunity to study early human neural development (21-23). We have developed highly reproducible protocols for controlled induction of differentiation of hESC toward a neural lineage, giving rise to enriched populations of proliferating, developmentally multipotent early NPC (24,25).Here, we have employed experimental HCMV infection in a dynamic model of controlled differentiation of hESC into neural precursors, to gain insight into the molecular events mediating HCMV infection during early human neurodevelopment.We first sought to track the earliest stage of neural differentiation at which the cells become susceptible to HCMV. To this end, differentiation of hESC was induced in floating cell clusters toward the formation of neural spheres (Fig. 1A) as detailed previously (24, 25). The emerging neural spheres were infected at sequential time points along the neural differentiation process with the broadly tropic HCMV TB40/E strain expressing green fluorescent protein (GFP) (26,27) (Fig. 1B). In accordance with previous studies (25), immunostaining analysis for the pluripotent stem cell marker TRA 1-81 and the neural differentiation marker PSA-NCAM showed that the majority of the cells underwent early neural differentiation within 11 days (Fig. 1C). Importantly, we have identified a transition toward HCMV susceptibility occurring between 4 and 11 days post-differentiation induction (Fig. 1B). A similar susceptibility pattern was observed for low-passage-number, cell-associated, clinic...

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“…However, permissiveness to virus replication is a characteristic that avian stem cells may not share with other species. In fact, human and murine pluripotent stem cells were non-permissive to infection with multiple viruses, such as hepatitis C virus (HCV) [45], varicella zoster virus (VZV) [46], and only moderately permissive to Influenza A virus and human Herpes-Simplex virus-1 [47]. At least for HCV and VZV, permissiveness to infection appeared to increase with the degree of cellular differentiation [45, 46].…”

Section: Discussionmentioning

confidence: 99%

“…In fact, human and murine pluripotent stem cells were non-permissive to infection with multiple viruses, such as hepatitis C virus (HCV) [45], varicella zoster virus (VZV) [46], and only moderately permissive to Influenza A virus and human Herpes-Simplex virus-1 [47]. At least for HCV and VZV, permissiveness to infection appeared to increase with the degree of cellular differentiation [45, 46]. This might suggest that, contrary to what observed with other viruses, NDV infection is not particularly restricted by the degree of cellular differentiation, or that chicken iPSCs have specific characteristics that are suitable for virus growth.…”

Section: Discussionmentioning

confidence: 99%

Derivation of chicken induced pluripotent stem cells tolerant to Newcastle disease virus-induced lysis through multiple rounds of infection

Susta

Hutcheson

et al. 2016

Virol J

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BackgroundNewcastle disease (ND), caused by Newcastle disease virus (NDV), is a devastating disease of poultry and wild birds. ND is prevented by rigorous biocontainment and vaccination. One potential approach to prevent spread of the virus is production of birds that show innate resistance to NDV-caused disease. Induced pluripotent stem cell (iPSC) technology allows adult cells to be reprogrammed into an embryonic stem cell-like state capable of contributing to live offspring and passing on unique traits in a number of species. Recently, iPSC approaches have been successfully applied to avian cells. If chicken induced pluripotent stem cells (ciPSCs) are genetically or epigenetically modified to resist NDV infection, it may be possible to generate ND resistant poultry. There is limited information on the potential of ciPSCs to be infected by NDV, or the capacity of these cells to become resistant to infection. The aim of the present work was to assess the characteristics of the interaction between NDV and ciPSCs, and to develop a selection method that would increase tolerance of these cells to NDV-induced cellular damage.ResultsResults showed that ciPSCs were permissive to infection with NDV, and susceptible to virus-mediated cell death. Since ciPSCs that survived infection demonstrated the ability to recover quickly, we devised a system to select surviving cells through multiple infection rounds with NDV. ciPSCs that sustained 9 consecutive infections had a statistically significant increase in survival (up to 36 times) compared to never-infected ciPSCs upon NDV infection (tolerant cells). Increased survival was not caused by a loss of permissiveness to NDV replication. RNA sequencing followed by enrichment pathway analysis showed that numerous metabolic pathways where differentially regulated between tolerant and never-infected ciPSCs.ConclusionsResults demonstrate that ciPSCs are permissive to NDV infection and become increasingly tolerant to NDV under selective pressure, indicating that this system could be applied to study mechanisms of cellular tolerance to NDV.Electronic supplementary materialThe online version of this article (doi:10.1186/s12985-016-0659-3) contains supplementary material, which is available to authorized users.

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Varicella-Zoster Virus Infects Human Embryonic Stem Cell-Derived Neurons and Neurospheres but Not Pluripotent Embryonic Stem Cells or Early Progenitors

Cited by 35 publications

References 28 publications

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

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

Transition toward Human Cytomegalovirus Susceptibility in Early Human Embryonic Stem Cell-Derived Neural Precursors

Derivation of chicken induced pluripotent stem cells tolerant to Newcastle disease virus-induced lysis through multiple rounds of infection

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