Varicella-zoster virus (VZV) initiates primary infection by upper respiratory mucosal inoculation, causing varicella. VZV persists in sensory ganglia and reactivation from latency may result in zoster. The VZV genome has at least 71 known or predicted open reading frames (ORFs) but understanding how these gene products function in virulence is difficult because VZV is highly human-specific. We have addressed this obstacle by investigating VZV infection of human skin, T cell and dorsal root ganglia xenografts in the severe combined immunodeficiency (SCID) mouse model. Together with clinical observations, this work supports a new paradigm of VZV pathogenesis that incorporates a critical role for T cells as a vehicle for VZV delivery to both skin and neuronal sites of latency. VZV infects human tonsil T cells very efficiently and infected tonsil T cells transfer VZV into skin xenografts, where lesion formation occurs gradually against a barrier of a potent innate epidermal cell response. VZV-infected T cells also exit from the circulation into DRG xenografts, indicating that VZV can reach neurons by this mechanism. T cell tropism is an important difference between the life cycles of VZV and herpes simplex viruses 1 and 2. We propose ?state of the art? screening methods to show how takeover of T cells by VZV modifies cell protein synthesis and activation and alters T cell gene regulation in combination with experiments to define how specific VZV gene products function to achieve these changes in the infected T cell. We will investigate VZV effects on T cell signaling pathways, T cell protein synthesis and T cell gene expression at single cell level using validated screening techniques that have been applied to studies of human T cell biology;methods will include multiparameter flow cytometric analysis of phosphoproteins, multiplex cytokine/chemokine assays and microfluidics-based profiling of gene expression (Aim 1). Based on our preliminary observations, we will examine the contributions of three VZV proteins, immediate early 4 (IE4), immediate early 62 (IE62) and ORF3 protein to VZV T cell tropism (Aim 2). Screening experiments that identify major VZV-induced consequences for the T cell will set the stage for additional studies of the mechanisms by which the virus achieves these effects. VZV is likely to target some pathways common to other T cell tropic viruses and strategies for applying new technologies to investigate viral pathogenesis should emerge. From a public health perspective, this work should provide information to improve the live attenuated Oka strain vaccines for varicella and zoster. While these vaccines are safe and beneficial, this work will be relevant for developing a ?2nd generation? VZV vaccine which consists of a VZV recombinant virus that has impaired T cell tropism and therefore less potential to disseminate in high risk patients and to establish latency in healthy individuals.

Public Health Relevance

Chickenpox and herpes zoster (shingles) are caused by varicella-zoster virus (VZV). These infections remain an important public health problem in the United States. The licensed VZV vaccines induce protection in most children and elderly adults but complications can occur in healthy vaccine recipients and in those who have diseases that impair their immune systems. Our goal is to identify strategies for creating better vaccines to prevent VZV infections in healthy and high risk people.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI020459-31
Application #
8636967
Study Section
Special Emphasis Panel (ZRG1-IDM-B (03))
Program Officer
Challberg, Mark D
Project Start
1994-08-01
Project End
2015-04-30
Budget Start
2014-05-01
Budget End
2015-04-30
Support Year
31
Fiscal Year
2014
Total Cost
$438,199
Indirect Cost
$144,223
Name
Stanford University
Department
Pediatrics
Type
Schools of Medicine
DUNS #
009214214
City
Stanford
State
CA
Country
United States
Zip Code
94305
Zerboni, Leigh; Sung, Phillip; Sommer, Marvin et al. (2018) The C-terminus of varicella-zoster virus glycoprotein M contains trafficking motifs that mediate skin virulence in the SCID-human model of VZV pathogenesis. Virology 523:110-120
Zerboni, Leigh; Sung, Phillip; Lee, Gordon et al. (2018) Age-Associated Differences in Infection of Human Skin in the SCID Mouse Model of Varicella-Zoster Virus Pathogenesis. J Virol 92:
Sen, Nandini; Sung, Phillip; Panda, Arjun et al. (2018) Distinctive Roles for Type I and Type II Interferons and Interferon Regulatory Factors in the Host Cell Defense against Varicella-Zoster Virus. J Virol 92:
Sullivan, Nicole L; Reuter-Monslow, Morgan A; Sei, Janet et al. (2018) Breadth and Functionality of Varicella-Zoster Virus Glycoprotein-Specific Antibodies Identified after Zostavax Vaccination in Humans. J Virol 92:
Oliver, Stefan L; Yang, Edward; Arvin, Ann M (2017) Dysregulated Glycoprotein B-Mediated Cell-Cell Fusion Disrupts Varicella-Zoster Virus and Host Gene Transcription during Infection. J Virol 91:
Fran├žois, Sylvie; Sen, Nandini; Mitton, Bryan et al. (2016) Varicella-Zoster Virus Activates CREB, and Inhibition of the pCREB-p300/CBP Interaction Inhibits Viral Replication In Vitro and Skin Pathogenesis In Vivo. J Virol 90:8686-97
Khalil, Mohamed I; Che, Xibing; Sung, Phillip et al. (2016) Mutational analysis of varicella-zoster virus (VZV) immediate early protein (IE62) subdomains and their importance in viral replication. Virology 492:82-91
Oliver, Stefan L; Yang, Edward; Arvin, Ann M (2016) Varicella-Zoster Virus Glycoproteins: Entry, Replication, and Pathogenesis. Curr Clin Microbiol Rep 3:204-215
Sen, Nandini; Arvin, Ann M (2016) Dissecting the Molecular Mechanisms of the Tropism of Varicella-Zoster Virus for Human T Cells. J Virol 90:3284-7
Sen, Nandini; Mukherjee, Gourab; Arvin, Ann M (2015) Single cell mass cytometry reveals remodeling of human T cell phenotypes by varicella zoster virus. Methods 90:85-94

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