Reactivation of Latent Cytomegalovirus Infection in Mouse Brain Cells Detected after Transfer to Brain Slice Cultures

Tsutsui, Yoshihiro; Kawasaki, Hideya; Kosugi, Isao

doi:10.1128/jvi.76.14.7247-7254.2002

Cited by 40 publications

(30 citation statements)

References 62 publications

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“…Instead, the current data suggest that neural stem cells might constitute the site of latency in the CNS. Indeed, when brain slices from latently infected mice were cultured for extended periods of time, reactivated murine CMV IE expression was mainly detected in immature neural stem cells lining the ventricular cavities (41). Our observations that the HCMV genome persisted in individual pNSCs in vitro for an extended period of time support the idea that cells in the early stages of neural development might constitute a reservoir of virus in the CNS.…”

Section: Discussionsupporting

confidence: 71%

Human Cytomegalovirus Infection of Human Embryonic Stem Cell-Derived Primitive Neural Stem Cells Is Restricted at Several Steps but Leads to the Persistence of Viral DNA

Belzile

Stark

Yeo

et al. 2014

J Virol

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Congenital human cytomegalovirus (HCMV) infection is a major cause of central nervous system structural anomalies and sensory impairments. It is likely that the stage of fetal development, as well as the state of differentiation of susceptible cells at the time of infection, affects the severity of the disease. We used human embryonic stem (ES) cell-derived primitive prerosette neural stem cells (pNSCs) and neural progenitor cells (NPCs) maintained in chemically defined conditions to study HCMV replication in cells at the early stages of neural development. In contrast to what was observed previously using fetus-derived NPCs, infection of ES cell-derived pNSCs with HCMV was nonprogressive. At a low multiplicity of infection, we observed only a small percentage of cells expressing immediate-early genes (IE) and early genes. IE expression was found to be restricted to cells negative for the anterior marker FORSE-1, and treatment of pNSCs with retinoic acid restored IE expression. Differentiation of pNSCs into NPCs restored IE expression but not the transactivation of early genes. Virions produced in NPCs and pNSCs were exclusively cell associated and were mostly non-neural tropic. Finally, we found that viral genomes could persist in pNSC cultures for up to a month after infection despite the absence of detectable IE expression by immunofluorescence, and infectious virus could be produced upon differentiation of pNSCs to neurons. In conclusion, our results highlight the complex array of hurdles that HCMV must overcome in order to infect primitive neural stem cells and suggest that these cells might act as a reservoir for the virus. IMPORTANCEHuman cytomegalovirus (HCMV) is a betaherpesvirus that is highly prevalent in the population. HCMV infection is usually asymptomatic but can lead to severe consequences in immunosuppressed individuals. HCMV is also the most important infectious cause of congenital developmental birth defects. Manifestations of fetal HCMV disease range from deafness and learning disabilities to more severe symptoms such as microcephaly. In this study, we have used embryonic stem cells to generate primitive neural stem cells and have used these to model HCMV infection of the fetal central nervous system (CNS) in vitro. Our results reveal that these cells, which are similar to those present in the developing neural tube, do not support viral replication but instead likely constitute a viral reservoir. Future work will define the effect of viral persistence on cellular functions as well as the exogenous signals leading to the reactivation of viral replication in the CNS. Human cytomegalovirus (HCMV) is the most important infectious cause of congenital developmental birth defects, with the central nervous system (CNS) being a main target. The incidence of fetal transmission is between 1 and 4% of live births. At birth, 5 to 10% of in utero-infected infants display symptoms of CMV inclusion disease, which can range from mild to severe and is often lethal. Neural developmental abnormalities i...

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Section: Discussionsupporting

confidence: 71%

Human Cytomegalovirus Infection of Human Embryonic Stem Cell-Derived Primitive Neural Stem Cells Is Restricted at Several Steps but Leads to the Persistence of Viral DNA

Belzile

Stark

Yeo

et al. 2014

J Virol

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“…Furthermore, the clinical manifestations on some animal models are different from those observed in the human central nervous system (CNS) (Woolf et al 2007). Nevertheless, these animal models have provided evidence that the highest CMV susceptibility in CNS is presented by neural precursor cells (NPCs) (Tsutsui et al 2002). Infection of these cells might be the cause of microcephaly and mental retardation since it has been observed that murine CMV inhibits NPC proliferation and migration (Shinmura et al 1997;Tsutsui et al 2008).…”

Section: Introductionmentioning

confidence: 99%

Effects of cytomegalovirus infection in human neural precursor cells depend on their differentiation state

et al. 2015

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Cytomegalovirus (CMV) is the most common cause of congenital infection in developed countries and a major cause of neurological disability in children. Although CMV can affect multiple organs, the most important sequelae of intrauterine infection are related to lesions of the central nervous system. However, little is known about the pathogenesis and the cellular events responsible for neuronal damage in infants with congenital infection. Some studies have demonstrated that neural precursor cells (NPCs) show the greatest susceptibility to CMV infection in the developing brain. We sought to establish an in vitro model of CMV infection of the developing brain in order to analyze the cellular events associated with invasion by this virus. To this end, we employed two cell lines as a permanent source of NPC, avoiding the continuous use of human fetal tissue, the human SK-N-MC neuroblastoma cell line, and an immortalized cell line of human fetal neural origin, hNS-1. We also investigated the effect of the differentiation stage in relation to the susceptibility of these cell lines by comparing the neuroblastoma cell line with the multipotent cell line hNS-1. We found that the effects of the virus were more severe in the neuroblastoma cell line. Additionally, we induced hNS-1 to differentiate and evaluated the effect of CMV in these differentiated cells. Like SK-N-MC cells, hNS-1-differentiated cells were also susceptible to infection. Viability of differentiated hNS-1 cells decreased after CMV infection in contrast to undifferentiated cells. In addition, differentiated hNS-1 cells showed an extensive cytopathic effect whereas the effect was scarce in undifferentiated cells. We describe some of the effects of CMV in neural stem cells, and our observations suggest that the degree of differentiation is important in the acquisition of susceptibility.

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“…Murine CMV latency is not limited to cells of myeloid lineage (23,60), and the same might be true for HCMV. The periventricular location of neuronal-cell precursors from which murine CMV reactivates upon culturing of latently infected mouse brain explants (60) is reminiscent of HCMV's proclivity for subependymal neuronal cells in the ventriculoencephalitis that results from HCMV reactivation in persons with advanced AIDS (5,64).…”

mentioning

confidence: 99%

“…The periventricular location of neuronal-cell precursors from which murine CMV reactivates upon culturing of latently infected mouse brain explants (60) is reminiscent of HCMV's proclivity for subependymal neuronal cells in the ventriculoencephalitis that results from HCMV reactivation in persons with advanced AIDS (5,64). The mechanisms that underlie HCMV reactivation disease in the central nervous system are unknown, though terminal differentiation of the neuronal lineage is linked to permissiveness for HCMV replication (43).…”

mentioning

confidence: 99%

Reversal of Human Cytomegalovirus Major Immediate-Early Enhancer/Promoter Silencing in Quiescently Infected Cells via the Cyclic AMP Signaling Pathway

Keller¹,

Wu²,

Andrews

et al. 2007

J Virol

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Reactivation of Latent Cytomegalovirus Infection in Mouse Brain Cells Detected after Transfer to Brain Slice Cultures

Cited by 40 publications

References 62 publications

Human Cytomegalovirus Infection of Human Embryonic Stem Cell-Derived Primitive Neural Stem Cells Is Restricted at Several Steps but Leads to the Persistence of Viral DNA

Human Cytomegalovirus Infection of Human Embryonic Stem Cell-Derived Primitive Neural Stem Cells Is Restricted at Several Steps but Leads to the Persistence of Viral DNA

Effects of cytomegalovirus infection in human neural precursor cells depend on their differentiation state

Reversal of Human Cytomegalovirus Major Immediate-Early Enhancer/Promoter Silencing in Quiescently Infected Cells via the Cyclic AMP Signaling Pathway

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