Human embryonic stem cells and derived contractile embryoid bodies are susceptible to Coxsakievirus B infection and respond to interferon Iβ treatment

Scassa, María Élida; Giusti, C; Questa, María; Pretre, Gabriela; Richardson, Guillermo Agustín Videla; Blüguermann, Carolina; Romorini, Leonardo; Ferrer, María Florencia; Sevlever, Gustavo; Miriuka, Santiago Gabriel; Gómez, Ricardo Martín

doi:10.1016/j.scr.2010.09.002

Cited by 21 publications

(25 citation statements)

References 36 publications

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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).…”

mentioning

confidence: 86%

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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confidence: 86%

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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“…However, it has been reported that adenovirus does not effectively infect and replicate in some hESC lines and infection is correlated with the expression of coxsackievirus receptor (CAR) but not ␣ v -integrin in naïve hESC (2). Others have reported that the coxsackievirus infects several lines of undifferentiated hESC (21).…”

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confidence: 99%

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

Dukhovny

Sloutskin

Markus

et al. 2012

J Virol

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bPluripotent human stem cells are a powerful tool for the generation of differentiated cells that can be used for the study of human disease. We recently demonstrated that neurons derived from pluripotent human embryonic stem cells (hESC) can be infected by the highly host-restricted human alphaherpesvirus varicella-zoster virus (VZV), permitting the interaction of VZV with neurons to be readily evaluated in culture. In the present study, we examine whether pluripotent hESC and neural progenitors at intermediate stages of differentiation are permissive for VZV infection. We demonstrate here that VZV infection is blocked in naïve hESC. A block to VZV replication is also seen when a bacterial artificial chromosome (BAC) containing the VZV genome is transfected into hESC. In contrast, related alphaherpesviruses herpes simplex virus 1 (HSV-1) and pseudorabies virus (PrV) productively infect naïve hESC in a cell-free manner, and PrV replicates from a BAC transfected into hESC. Neurons differentiate from hESC via neural progenitor intermediates, as is the case in the embryo. The first in vitro stage at which permissiveness of hESC-derived neural precursors to VZV replication is observed is upon formation of "neurospheres," immediately after detachment from the inductive stromal feeder layer. These findings suggest that hESC may be useful in deciphering the yet enigmatic mechanisms of specificity of VZV infection and replication. V aricella-zoster virus (VZV) replication is highly host restricted, growing efficiently only in human cells. In varicella, VZV typically infects and replicates in cutaneous fibroblasts and epidermal cells as well as several types of immune cells. VZV infections of central nervous system (CNS) vasculature are also not uncommonly observed, the virus infecting smooth muscle actinexpressing cells in vessel walls (16). VZV infects effectively primarily in a cell-associated manner in vitro, and it is thought that cell-to-cell spread occurs in most tissues. Cell-free virus is made in vivo by keratinocytes and is present in cutaneous vesicles (8), and released VZV appears to be an important component of T cell-toskin transmission in vivo (reviewed in reference 1).VZV infection of neurons is essential for establishment of latency and the ability to reactivate to cause herpes zoster. Initial neuronal infection by VZV is via cutaneous axons and retrograde transport to peripheral somatic and autonomic ganglia and/or by infected circulating lymphocytes that infiltrate the ganglia (28). VZV replicates in both neurons and ganglionic support cells of somatic and cranial peripheral sensory ganglia both upon initial infection and upon reactivation. Importantly, VZV causes a plethora of CNS diseases (due at least in part to infection of the vasculature) (16) and ocular diseases (reviewed in reference 9). The growth of VZV in neurons and the interactions that govern latency and reactivation have proven to be difficult to study outside the human host because of the species restriction of infection and the limited...

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“…In this sense, it was reported that hESCs robustly express CVB receptors CAR and DAF and are susceptible to CVB3 infection. In particular, CAR transcripts were always more abundant in hESCs than in differentiated counterparts obtained using embryoid body Apoptosis (2012) 17:132-142 139 based hESCs-differentiation protocol [44]. Taken together, all these observations make the study of the mechanisms of cell death triggered by CVB3 virus infection in hESCs of growing interest.…”

Section: Discussionmentioning

confidence: 94%

“…On this sense, it was recently reported that non-transformed highly replicative human embryonic stem cell lines robustly express CAR and DAF receptors and are susceptible to CVB1-5 strains infection. Importantly, IFN-Ib treatment significantly reduced viral replication, suggesting that type I IFN signalling is also involved in the control of CVB infection in these cells [44]. hESCs emerged then as good candidates to study CVB3-induced cytopathic effects, and because of that we aim to determine the mechanisms of cell death triggered by CVB3 infection in hESCs.…”

Section: Introductionmentioning

confidence: 99%

Activation of apoptotic signalling events in human embryonic stem cells upon Coxsackievirus B3 infection

et al. 2011

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Human embryonic stem cells (hESCs) are self-renewing pluripotent cells that can differentiate to a wide range of specialized cells and hold great promise as models for human development and disease, as well as for drug discovery and cell-replacement therapies. Group B Coxsackie viruses (CVBs) produce acute myocarditis, pancreatitis, non-septic meningitis and encephalitis in neonates, children and young adults. Moreover, CVBs can produce spontaneous miscarriage after early embryo infection. It was reported that hESCs express CVBs receptors and are susceptible to CVB3 infection. Apoptosis is one of the hallmarks of CVBs infection although details regarding CVB3 involvement in the apoptotic processes remain elusive. In order to evaluate the mechanisms of cell death induced by CVB3 in these pluripotent cells, we infected HUES-5 (H5) and WA01 (H1) hESC lines with CVB3. After validating the maintenance of stemness in these hESC lines when grown as confluent monolayers in feeder-free conditions, we analysed several aspects of programmed cell death triggered by CVB3. In all cases, we detected chromatin condensation, DNA fragmentation and caspase-9 and 3 cleavages. Moreover, we observed the presence of cleaved PARP product which was preceded by the appearance of p17, the catalytically active fragment of caspase-3. Mitochondrial function assays revealed a MOI dependent decrease in cell viability at 24 h post-infection (pi). No appreciable modifications in Bcl-2, Bcl-X(L) and Bax protein levels were observed upon CVB3 infection during 5-24 h observation period. However, a marked decrease in pro-apoptotic Bad abundance was detected without changes in its mRNA levels. In this study we found that the hESCs are highly susceptible to CVB3 infection and display elevated apoptosis rates, thus emerging as suitable human non-transformed in vitro models to study CVB3-induced apoptosis and resulting relevant to understand CVBs pathogenesis.

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Human embryonic stem cells and derived contractile embryoid bodies are susceptible to Coxsakievirus B infection and respond to interferon Iβ treatment

Cited by 21 publications

References 36 publications

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

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

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

Activation of apoptotic signalling events in human embryonic stem cells upon Coxsackievirus B3 infection

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