scholarly journals Improved Identification and Differentiation of Varicella-Zoster Virus (VZV) Wild-Type Strains and an Attenuated Varicella Vaccine Strain Using a VZV Open Reading Frame 62-Based PCR

2000 ◽  
Vol 38 (9) ◽  
pp. 3156-3160 ◽  
Author(s):  
Vladimir N. Loparev ◽  
Takele Argaw ◽  
Philip R. Krause ◽  
Michiko Takayama ◽  
D. Scott Schmid
Keyword(s):  
Varicella Zoster Virus ◽  
Vaccine Strain ◽  
Varicella Vaccine ◽  
Open Reading Frame ◽  
Cleavage Sites ◽  
Wild Type ◽  
Reading Frame ◽  
Varicella Zoster ◽  
Enzyme Cleavage ◽  
Type Strains

A new method was developed to identify and differentiate varicella-zoster virus (VZV) wild-type strains from the attenuated varicella Oka vaccine strain. The PCR technique was used to amplify a VZV open reading frame (ORF) 62 region. A single specific amplicon of 268 bp was obtained from 71 VZV clinical isolates and several laboratory strains. Subsequent digestion of the VZV ORF 62 amplicons with SmaI enabled accurate strain differentiation (threeSmaI sites were present in amplicons of vaccine strain VZV, compared with two enzyme cleavage sites for all other VZV strains tested). This method accurately differentiated the Oka vaccine strain from wild-type VZV strains circulating in countries representing all six populated continents. Moreover, the assay more reliably distinguished wild-type Japanese strains from the vaccine strain than did previously described methods.

scholarly journals Open Reading Frame S/L of Varicella-Zoster Virus Encodes a Cytoplasmic Protein Expressed in Infected Cells

2000 ◽  
Vol 74 (23) ◽  
pp. 11311-11321 ◽  
Author(s):  
George W. Kemble ◽  
Paula Annunziato ◽  
Octavian Lungu ◽  
Ruth E. Winter ◽  
Tai-An Cha ◽  
...  
Keyword(s):  
Varicella Zoster Virus ◽  
Skin Lesions ◽  
Open Reading Frame ◽  
Plaque Morphology ◽  
Virus Reactivation ◽  
Wild Type ◽  
Reading Frame ◽  
Varicella Zoster ◽  
L Protein ◽  
Infected Cells

ABSTRACT We report the discovery of a novel gene in the varicella-zoster virus (VZV) genome, designated open reading frame (ORF) S/L. This gene, located at the left end of the prototype VZV genome isomer, expresses a polyadenylated mRNA containing a splice within the 3′ untranslated region in virus-infected cells. Sequence analysis reveals significant differences between the ORF S/Ls of wild-type and attenuated strains of VZV. Antisera raised to a bacterially expressed portion of ORF S/L reacted specifically with a 21-kDa protein synthesized in cells infected with a VZV clinical isolate and with the original vaccine strain of VZV (Oka-ATCC). Cells infected with other VZV strains, including a wild-type strain that has been extensively passaged in tissue culture and commercially produced vaccine strains of Oka, synthesize a family of proteins ranging in size from 21 to 30 kDa that react with the anti-ORF S/L antiserum. MeWO cells infected with recombinant VZV harboring mutations in the C-terminal region of the ORF S/L gene lost adherence to the stratum and adjacent cells, resulting in an altered plaque morphology. Immunohistochemical analysis of VZV-infected cells demonstrated that ORF S/L protein localizes to the cytoplasm. ORF S/L protein was present in skin lesions of individuals with primary or reactivated infection and in the neurons of a dorsal root ganglion during virus reactivation.

scholarly journals Varicella Vaccine Meningitis as a Complication of Herpes Zoster in Twice-Immunized Immunocompetent Adolescents

2020 ◽  
Vol 35 (13) ◽  
pp. 889-895 ◽  
Author(s):  
Veena Ramachandran ◽  
Stephen C. Elliott ◽  
Kathie L. Rogers ◽  
Randall J. Cohrs ◽  
Miles Weinberger ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Herpes Zoster ◽  
Varicella Zoster Virus ◽  
Varicella Vaccine ◽  
The United States ◽  
Vaccine Virus ◽  
Virus Reactivation ◽  
Wild Type ◽  
Varicella Zoster ◽  
Intravenous Acyclovir

Varicella-zoster virus vaccination is recommended for virtually all young children in the United States, Canada, and several other countries. Varicella vaccine is a live attenuated virus that retains some of its neurotropic properties. Herpes zoster caused by vaccine virus still occurs in immunized children, although the rate is much lower than in children who had wild-type varicella. It was commonly thought that 2 varicella vaccinations would protect children against the most serious complication of meningitis following herpes zoster; however, 2 meningitis cases have already been published. We now report a third case of varicella vaccine meningitis and define risk factors shared by all 3 immunized adolescents. The diagnosis in cerebrospinal fluid in this third case was verified by amplifying and sequencing portions of the viral genome, to document fixed alleles found only in the vaccine strain. Viral antibody was also detected in the cerebrospinal fluid by confocal microscopy. When compared with the other 2 cases, remarkably all 3 were 14 years old when meningitis occurred. All 3 were treated with intravenous acyclovir, with complete recovery. The adolescent in our case report also had recurrent asthma, which was treated with both prednisone tablets and beclomethasone inhaler before onset of meningitis. When the 3 cases were considered together, they suggested that immunity to varicella-zoster virus may be waning sufficiently in some twice-immunized adolescents to make them vulnerable to varicella vaccine virus reactivation and subsequent meningitis. This complication rarely happens in children after wild-type varicella.

Varicella-Zoster Virus (VZV) Virion-Associated Transactivator Open Reading Frame 62 Protein Enhances the Infectivity of VZV DNA

Virology ◽  
1994 ◽  
Vol 200 (1) ◽  
pp. 297-300 ◽  
Author(s):  
Masako Moriuchi ◽  
Hiroyuki Moriuchi ◽  
Stephen E. Straus ◽  
Jeffrey I. Cohen
Keyword(s):  
Varicella Zoster Virus ◽  
Open Reading Frame ◽  
Reading Frame ◽  
Varicella Zoster

Genotypes of varicella-zoster virus wild-type strains in Germany

2008 ◽  
Vol 80 (6) ◽  
pp. 1123-1130 ◽  
Author(s):  
A. Sauerbrei ◽  
R. Zell ◽  
A. Philipps ◽  
P. Wutzler
Keyword(s):  
Varicella Zoster Virus ◽  
Wild Type ◽  
Varicella Zoster ◽  
Type Strains

scholarly journals Nuclear Accumulation of IE62, the Varicella-Zoster Virus (VZV) Major Transcriptional Regulatory Protein, Is Inhibited by Phosphorylation Mediated by the VZV Open Reading Frame 66 Protein Kinase

2000 ◽  
Vol 74 (5) ◽  
pp. 2265-2277 ◽  
Author(s):  
Paul R. Kinchington ◽  
Karen Fite ◽  
Stephanie E. Turse
Keyword(s):  
Protein Kinase ◽  
Varicella Zoster Virus ◽  
Nuclear Localization ◽  
Kinase Activity ◽  
Open Reading Frame ◽  
Nuclear Accumulation ◽  
Protein Kinase Activity ◽  
Reading Frame ◽  
Varicella Zoster ◽  
In Cells

ABSTRACT IE62, the major transcriptional activator protein encoded by varicella-zoster virus (VZV), locates to the nucleus when expressed in transfected cells. We show here that cytoplasmic forms of IE62 accumulate in transfected and VZV-infected cells as the result of the protein kinase activity associated with VZV open reading frame 66 (ORF66). Expression of the ORF66 protein kinase but not the VZV ORF47 protein kinase impaired the ability of coexpressed IE62 to transactivate promoter-reporter constructs. IE62 that was coexpressed with the ORF66 protein accumulated predominantly in the cytoplasm, whereas the normal nuclear localization of other proteins was not affected by the ORF66 protein. In cells infected with VZV, IE62 accumulated in the cytoplasm at late times of infection, whereas in cells infected with a VZV recombinant unable to express ORF66 protein (ROka66S), IE62 was completely nuclear. Point mutations introduced into the predicted serine/threonine catalytic domain and ATP binding domain of ORF66 abrogated its ability to influence IE62 nuclear localization, indicating that the protein kinase activity was required. The region of IE62 that was targeted by ORF66 was mapped to amino acids 602 to 733. IE62 peptides containing this region were specifically phosphorylated in cells coexpressing the ORF66 protein kinase and in cells infected with wild-type VZV but were not phosphorylated in cells infected with ROka66S. We conclude that the ORF66 protein kinase phosphorylates IE62 to induce its cytoplasmic accumulation, most likely by inhibiting IE62 nuclear import.

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