5 results on '"Embryonic Stem Cells virology"'
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2. 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.
- Author
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Belzile JP, Stark TJ, Yeo GW, and Spector DH
- Subjects
- Cell Differentiation, Cytomegalovirus genetics, Cytomegalovirus Infections physiopathology, DNA, Viral genetics, Embryonic Stem Cells cytology, Female, Humans, Neural Stem Cells cytology, Virus Replication, Cytomegalovirus physiology, Cytomegalovirus Infections virology, DNA, Viral metabolism, Embryonic Stem Cells virology, Neural Stem Cells virology
- Abstract
Unlabelled: 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., Importance: Human 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.
- Published
- 2014
- Full Text
- View/download PDF
3. Human embryonic stem cell lines model experimental human cytomegalovirus latency.
- Author
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Penkert RR and Kalejta RF
- Subjects
- Adaptor Proteins, Signal Transducing metabolism, Antigens, CD34 analysis, Cell Culture Techniques methods, Chromatin metabolism, Co-Repressor Proteins, Gene Silencing, Histone Deacetylases metabolism, Humans, Molecular Chaperones, Nuclear Proteins metabolism, Virology methods, Cytomegalovirus physiology, Embryonic Stem Cells virology, Host-Pathogen Interactions, Virus Latency
- Abstract
Herpesviruses are highly successful pathogens that persist for the lifetime of their hosts primarily because of their ability to establish and maintain latent infections from which the virus is capable of productively reactivating. Human cytomegalovirus (HCMV), a betaherpesvirus, establishes latency in CD34(+) hematopoietic progenitor cells during natural infections in the body. Experimental infection of CD34(+) cells ex vivo has demonstrated that expression of the viral gene products that drive productive infection is silenced by an intrinsic immune defense mediated by Daxx and histone deacetylases through heterochromatinization of the viral genome during the establishment of latency. Additional mechanistic details about the establishment, let alone maintenance and reactivation, of HCMV latency remain scarce. This is partly due to the technical challenges of CD34(+) cell culture, most notably, the difficulty in preventing spontaneous differentiation that drives reactivation and renders them permissive for productive infection. Here we demonstrate that HCMV can establish, maintain, and reactivate in vitro from experimental latency in cultures of human embryonic stem cells (ESCs), for which spurious differentiation can be prevented or controlled. Furthermore, we show that known molecular aspects of HCMV latency are faithfully recapitulated in these cells. In total, we present ESCs as a novel, tractable model for studies of HCMV latency.
- Published
- 2013
- Full Text
- View/download PDF
4. Varicella-zoster virus infects human embryonic stem cell-derived neurons and neurospheres but not pluripotent embryonic stem cells or early progenitors.
- Author
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Dukhovny A, Sloutskin A, Markus A, Yee MB, Kinchington PR, and Goldstein RS
- Subjects
- Cell Differentiation, Cells, Cultured, Embryonic Stem Cells cytology, Herpesvirus 3, Human genetics, Humans, Neurons cytology, Pluripotent Stem Cells cytology, Virus Replication, Embryonic Stem Cells virology, Herpesvirus 3, Human physiology, Neurons virology, Pluripotent Stem Cells virology
- Abstract
Pluripotent 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.
- Published
- 2012
- Full Text
- View/download PDF
5. Primer binding site-dependent restriction of murine leukemia virus requires HP1 binding by TRIM28.
- Author
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Wolf D, Cammas F, Losson R, and Goff SP
- Subjects
- Animals, Binding Sites, Cell Line, Chromobox Protein Homolog 5, Embryonal Carcinoma Stem Cells virology, Embryonic Stem Cells virology, Mice, Nuclear Proteins metabolism, Point Mutation, Protein Binding genetics, Transcription Factors metabolism, Tripartite Motif-Containing Protein 28, Chromosomal Proteins, Non-Histone metabolism, Leukemia Virus, Murine physiology, Repressor Proteins metabolism, Virus Replication
- Abstract
TRIM28 is a transcriptional corepressor which is required for primer binding site (PBS)-dependent restriction of murine leukemia virus (MLV) replication in embryonic stem and embryonic carcinoma (EC) cells. PBS-dependent restriction of MLV leads to transcriptional silencing of the integrated provirus and has been shown to correlate with TRIM28-mediated recruitment of HP1 to the silenced loci. Here we show, using a cell line with a point mutation in the HP1 binding domain of TRIM28, that interaction with HP1 is absolutely required for the PBS-dependent restriction of MLV in the F9 EC cell line.
- Published
- 2008
- Full Text
- View/download PDF
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