Productive Varicella-Zoster Virus Infection of Cultured Intact Human Ganglia

ABSTRACT Varicella-zoster virus (VZV) is a species-specific herpesvirus which infects sensory ganglia. We have developed a model of infection of human intact explant dorsal root ganglia (DRG). Following exposure of DRG to VZV, viral antigens were detected in neurons and nonneuronal cells. Enveloped virions were visualized by transmission electron microscopy in neurons and nonneuronal cells and within the extracellular space. Moreover, rather than remaining highly cell associated during infection of cultured cells, such as fibroblasts, cell-free VZV was released from infected DRG. This model enables VZV infection of ganglionic cells to be studied in the context of intact DRG.

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Upregulation of CXCL10 in Human Dorsal Root Ganglia during Experimental and Natural Varicella-Zoster Virus Infection

Journal of Virology ◽

10.1128/jvi.01816-10 ◽

2010 ◽

Vol 85 (1) ◽

pp. 626-631 ◽

Cited By ~ 33

Author(s):

M. Steain ◽

K. Gowrishankar ◽

M. Rodriguez ◽

B. Slobedman ◽

A. Abendroth

Keyword(s):

Virus Infection ◽

Varicella Zoster Virus ◽

Dorsal Root Ganglia ◽

Dorsal Root ◽

Varicella Zoster Virus Infection ◽

Varicella Zoster

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Varicella-Zoster Virus Infection of Human Foreskin Fibroblast Cells Results in Atypical Cyclin Expression and Cyclin-Dependent Kinase Activity

Journal of Virology ◽

10.1128/jvi.00163-06 ◽

2006 ◽

Vol 80 (11) ◽

pp. 5577-5587 ◽

Cited By ~ 17

Author(s):

Stacey A. Leisenfelder ◽

Jennifer F. Moffat

Keyword(s):

Cell Cycle ◽

Varicella Zoster Virus ◽

Cultured Cells ◽

Cyclin B1 ◽

Human Infection ◽

Dermal Fibroblasts ◽

Cyclin D3 ◽

Varicella Zoster Virus Infection ◽

Varicella Zoster ◽

Protein Levels

ABSTRACT In its course of human infection, varicella-zoster virus (VZV) infects rarely dividing cells such as dermal fibroblasts, differentiated keratinocytes, mature T cells, and neurons, none of which are actively synthesizing DNA; however, VZV is able to productively infect them and use their machinery to replicate the viral genome. We hypothesized that VZV alters the intracellular environment to favor viral replication by dysregulating cell cycle proteins and kinases. Cyclin-dependent kinases (CDKs) and cyclins displayed a highly unusual profile in VZV-infected confluent fibroblasts: total amounts of CDK1, CDK2, cyclin B1, cyclin D3, and cyclin A protein increased, and kinase activities of CDK2, CDK4, and cyclin B1 were strongly and simultaneously induced. Cyclins B1 and D3 increased as early as 24 h after infection, concurrent with VZV protein synthesis. Confocal microscopy indicated that cyclin D3 overexpression was limited to areas of IE62 production, whereas cyclin B1 expression was irregular across the VZV plaque. Downstream substrates of CDKs, including pRb, p107, and GM130, did not show phosphorylation by immunoblotting, and p21 and p27 protein levels were increased following infection. Finally, although the complement of cyclin expression and high CDK activity indicated a progression through the S and G2 phases of the cell cycle, DNA staining and flow cytometry indicated a possible G1/S blockade in infected cells. These data support earlier studies showing that pharmacological CDK inhibitors can inhibit VZV replication in cultured cells.

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Differential Requirement for Cell Fusion and Virion Formation in the Pathogenesis of Varicella-Zoster Virus Infection in Skin and T Cells

Journal of Virology ◽

10.1128/jvi.78.23.13293-13305.2004 ◽

2004 ◽

Vol 78 (23) ◽

pp. 13293-13305 ◽

Cited By ~ 29

Author(s):

Jaya Besser ◽

Minako Ikoma ◽

Konstanze Fabel ◽

Marvin H. Sommer ◽

Leigh Zerboni ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Varicella Zoster Virus ◽

Cell Fusion ◽

Cultured Cells ◽

Varicella Zoster Virus Infection ◽

Virion Assembly ◽

Varicella Zoster ◽

Human T Cell ◽

Virion Formation

ABSTRACT The protein product of varicella-zoster virus (VZV) ORF47 is a serine/threonine protein kinase and tegument component. Evaluation of two recombinants of the Oka strain, rOka47ΔC, with a C-terminal truncation of ORF47, and rOka47D-N, with a point mutation in the conserved kinase motif, showed that ORF47 kinase function was necessary for optimal VZV replication in human skin xenografts in SCID mice but not in cultured cells. We now demonstrate that rOka47ΔC and rOka47D-N mutants do not infect human T-cell xenografts. Differences in the growth of kinase-defective ORF47 mutants allowed an examination of requirements for VZV pathogenesis in skin and T cells in vivo. Although virion assembly was reduced and no virion transport to cell surfaces was observed, epidermal cell fusion persisted, and VZV polykaryocytes were generated by rOka47ΔC and rOka47D-N in skin. Virion assembly was also impaired in vitro, but VZV-induced cell fusion continued to cause syncytia in cultured cells infected with rOka47ΔC or rOka47D-N. Intracellular trafficking of envelope glycoprotein E and the ORF47 and IE62 proteins, components of the tegument, was aberrant without ORF47 kinase activity. In summary, normal VZV virion assembly appears to require ORF47 kinase function. Cell fusion was induced by ORF47 mutants in skin, and cell-cell spread occurred even though virion formation was deficient. VZV-infected T cells do not undergo cell fusion, and impaired virion assembly by ORF47 mutants was associated with a complete elimination of T-cell infectivity. These observations suggest a differential requirement for cell fusion and virion formation in the pathogenesis of VZV infection in skin and T cells.

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Varicella-Zoster Virus Infection Induces Autophagy in both Cultured Cells and Human Skin Vesicles

Journal of Virology ◽

10.1128/jvi.02670-08 ◽

2009 ◽

Vol 83 (11) ◽

pp. 5466-5476 ◽

Cited By ~ 50

Author(s):

Marie-Noëlle Takahashi ◽

Wallen Jackson ◽

Donna T. Laird ◽

Timothy D. Culp ◽

Charles Grose ◽

...

Keyword(s):

Varicella Zoster Virus ◽

Human Skin ◽

Cultured Cells ◽

Normal Skin ◽

Viral Gene ◽

Varicella Zoster Virus Infection ◽

Specific Cell ◽

Late Time ◽

Specific Staining ◽

Varicella Zoster

ABSTRACT When grown in cultured cells, varicella-zoster virus (VZV) forms many aberrant light particles and produces low titers. Various studies have explored the reasons for such a phenotype and have pointed to impaired expression of specific late genes and at lysosomal targeting of egressing virions as possible causes. In the studies presented here, we report that the autophagic degradation pathway was induced at late time points after VZV infection of cultured cells, as documented by immunoblot analysis of the cellular proteins LC3B and p62/SQSTM1, along with electron microscopy analysis, which demonstrated the presence of both early autophagosomes and late autophagic compartments. Autophagy was induced in infected cells even in the presence of phosphonoacetic acid, an inhibitor of viral late gene expression, thus suggesting that accumulation of immediate-early and early viral gene products might be the major stimulus for its induction. We also showed that the autophagic response was not dependent on a specific cell substrate, virus strain, or type of inoculum. Finally, using immunofluorescence imaging, we demonstrated autophagosome-specific staining in human zoster vesicles but not in normal skin. Thus, our results document that this innate immune response pathway is a component of the VZV infectious cycle in both cultured cells and the human skin vesicle, the final site of virion formation in the infected human host.

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