Competitive polymerase chain reaction was used to quantitate latent varicella-zoster virus (VZV) DNA in human trigeminal ganglia. Ganglionic DNA from five subjects was amplified with oligonucleotide primers specific for VZV gene 28. Two of the samples were also analyzed with primers specific for VZV gene 62. Our results indicated that there are 6 to 31 copies of the VZV genome in every 100,000 ganglionic cells.

Adult↗

PubMed1993

Treatment for glaucoma: adherence by the elderly.

OBJECTIVES: The purpose of this study was to determine the extent of nonadherence to treatment for glaucoma among elderly patients. METHODS: This was a retrospective cohort study of 2440 patients older than age 65 who were enrolled in the New Jersey Medicaid Program and who were newly initiated on a topical agent for the treatment of glaucoma. Two patient-specific measures of nonadherence were employed: (1) no filled prescription for any glaucoma medication over a 12-month period after the initiation of therapy and (2) number of days without therapy for glaucoma during this 12-month period. RESULTS: By the first measure, 569 patients (23%) were found to be nonadherent. The mean number of days without therapy during the study year was 112. Factors associated with nonadherence included the use of glaucoma medication requiring more than 2 administrations per day and the presence of multiple other medications in the patient's drug regimen. Patients started on multiple glaucoma medication were more adherent than those started on a single agent. Age and sex were not found to be predictors of nonadherence. CONCLUSIONS: Substantial nonadherence was found to be common in this population. More attention to the issue of nonadherence could result in important benefits in the preservation of sight.

Aged↗

PubMed1991

Simian varicella virus DNA in dorsal root ganglia.

Clinical, pathological, immunological, and virological evidence suggests that simian varicella virus (SVV) infection of primates is the counterpart of varicella-zoster virus infection of humans. To determine whether these two viruses share similarities in their properties during latency, we analyzed ganglia and brain of an African green monkey experimentally infected with SVV for the presence of viral nucleic acid using the polymerase chain reaction technique. We detected SVV DNA in dorsal root ganglia but not in brain of this monkey, which demonstrated no apparent clinical signs of SVV infection. Our results suggest that SVV becomes latent in monkey ganglia and that latency can develop in the absence of clinical varicella (chickenpox). These studies provide an animal model system to study varicella virus latency.

Animals↗

PubMed1991

Fatal varicella-zoster virus meningoradiculitis without skin involvement.

A 77-year-old man with T-cell lymphoma developed an acute fatal meningoradiculitis of cranial nerve roots and cauda equina, pathologically and virologically confirmed to be caused by varicella-zoster virus. This is the first report of fatal varicella-zoster virus-induced neurological disease in the absence of skin lesions. Varicella-zoster virus should be included in the differential diagnosis of acute radiculoneuropathy in the immunocompromised patient, particularly because antiviral treatment for varicella-zoster virus exists.

Aged↗

PubMed1991

Varicella-zoster virus-specific immunity after herpes zoster.

The frequency of varicella-zoster virus (VZV)-specific T lymphocytes was higher in elderly subjects who had herpes zoster infections than in age-matched controls. This increase in T cell response persisted for at least 2 years while antibody levels to VZV returned to control values at this time. There were no differences in T cell or antibody responses to VZV between individuals with and without postherpetic neuralgia. Elderly subjects who had not had herpes zoster had a comparable increase in VZV-specific T responder cell frequency after immunization with Oka strain VZV. The data suggest that the potential for a boost in T cell response to VZV persists in the elderly, and that immunization can elicit a T cell response in this age group.

Adult↗

PubMed1990

Latent varicella-zoster viral DNA in human trigeminal and thoracic ganglia.

BACKGROUND: Some human herpesviruses become latent in dorsal-root ganglia. Primary infection with the varicella-zoster virus causes chickenpox, followed by latency, and subsequent reactivation leading to shingles (zoster), but the frequency and distribution of latent virus have not been established. METHODS: Using the polymerase chain reaction, we performed postmortem examinations of trigeminal and thoracic ganglia of 23 subjects 33 to 88 years old who had not recently had chickenpox or shingles to identify the presence of latent varicella-zoster viral DNA. Oligonucleotide primers representing the origin of replication of the varicella-zoster virus and varicella-zoster virus gene 29 were used for amplification. RESULTS: Among the 22 subjects seropositive for the antibody to the virus, both the viral origin-of-replication and gene-29 sequences were detected in 13 of 15 subjects (87 percent) in whom trigeminal ganglia were examined and in 9 of 17 (53 percent) in whom thoracic ganglia were examined. Viral DNA was not detected in brain or mononuclear cells from the seropositive subjects. None of three thoracic ganglia from the one seronegative subject contained varicella-zoster viral DNA. CONCLUSIONS: These findings indicate that after primary infection with varicella-zoster virus (varicella), the virus becomes latent in many ganglia--more often in the trigeminal ganglia than in any thoracic ganglion--and that more than one region of the viral genome is present during latency.

Adult↗

PubMed1990

In-vitro synthesis of functional varicella zoster and herpes simplex viral thymidine kinase.

The varicella zoster virus (VZV) and herpes simplex virus type 1 (HSV-1) thymidine kinase (TK) genes were cloned into the transcription vector pGEM4. In-vitro translation (ivt) of RNA transcribed from these genes showed prominent expression of functional TK proteins with the expected molecular weights of 36 kD for VZV and 43, 39, and 38 kD for HSV-1. The TK proteins were recognized by rabbit anti-VZV and anti-HSV-1 antibodies, respectively. Analysis of the ivt products by thin-layer chromatography revealed the conversion of thymidine to its phosphorylated forms (TMP, TDP, and TTP) by both the VZV and HSV-1 TK genes. The estimated specific activities of the in-vitro translated VZV and HSV-1 TKs were comparable. VZV TK templates were linearized at internal restriction sites and RNAs transcribed from these templates directed the synthesis of polypeptides with sizes consistent with the colinearity of the VZV TK gene. Deletion of 107 amino acids at the carboxy terminus of the VZV TK gene abolished the in-vitro TK activity. In addition, immunoprecipitation of truncated proteins synthesized in vitro suggested the possible involvement of the region between amino acid residues 101 and 168 from the amino terminus of the VZV TK molecule in the formation of structures necessary for antigenicity.

Amino Acid Sequence↗

PubMed1988

Regulation of varicella zoster virus gene 27 translation in vitro by upstream sequences.

Northern blot analysis revealed the presence of varicella-zoster virus (VZV) gene 27 transcripts in infected cells. The Sal I-G DNA fragment, located in the unique long segment of the VZV genome and containing overlapping genes 26 and 27, was analyzed in an in vitro transcription-translation system. Translation of RNA transcribed from these open reading frames showed prominent expression of gene 27. Four different subclones were constructed to contain gene 27 with and without 100 base pairs (bp) of upstream sequences. Translation of RNA from these constructs using wheat germ extract or rabbit reticulocyte lysate indicated that the sequences upstream from the predicted initiation codon (AUG) of gene 27 downregulated the expression of this gene at the level of translation and that the predicted AUG within gene 27 was preferentially used.

Animals↗

PubMed1988

Expression of varicella-zoster virus glycoprotein I in cells infected with a vaccinia virus recombinant.

BSC-1 cells infected with a vaccinia virus recombinant containing the coding sequences for varicella-zoster virus (VZV) glycoprotein I (gpI) were analyzed by indirect immunofluorescence and immunoprecipitation for the expression and processing of gpI. The processing of gpI in cells infected with recombinant virus was the same as that observed during VZV infection. Immunofluorescence revealed localization of gpI to the membranes of recombinant virus-infected cells.

Cell Line↗

PubMed1988

Recognition of similar epitopes on varicella-zoster virus gpI and gpIV by monoclonal antibodies.

Two monoclonal antibodies, MAb43.2 and MAb79.0, prepared against varicella-zoster virus (VZV) proteins were selected to analyze VZV gpIV and gpI, respectively. MAb43.2 reacted only with cytoplasmic antigens, whereas MAb79.0 recognized both cytoplasmic and membrane antigens in VZV-infected cells. Immunoprecipitation of in vitro translation products with MAb43.2 revealed only proteins encoded by the gpIV gene, whereas MAb79.0 precipitated proteins encoded by the gpIV and gpI genes. Pulse-chase analysis followed by immunoprecipitation of VZV-infected cells indicated reactivity of MAb43.2 with three phosphorylated precursor species of gpIV and reactivity of MAb79.0 with the precursor and mature forms of gpI and gpIV. These results indicated that (i) MAb43.2 and MAb79.0 recognize different epitopes on VZV gpIV, (ii) glycosylation of gpIV ablates recognition by MAb43.2, and (iii) gpIV is phosphorylated. To map the binding site of MAb79.0 on gpI, the pGEM transcription vector, containing the coding region of the gpI gene, was linearized, and three truncated gpI DNA fragments were generated. RNA was transcribed from each truncated fragment by using SP6 RNA polymerase, translated in vitro in a rabbit reticulocyte lysate, and immunoprecipitated with MAb79.0 and human sera. The results revealed the existence of an antibody-binding site within 14 amino acid residues located between residues 109 to 123 on the predicted amino acid sequences of gpI. From the predicted amino acid sequences, 14 residues on gpI (residues 107 to 121) displayed a degree of similarity (36%) to two regions (residues 55 to 69 and 245 to 259) of gp IV. Such similarities may account for the binding of MAb79.0 to both VZV gpI and gpIV.

Amino Acid Sequence↗

PubMed1987

Induction of antibody against in vitro translation products encoded by varicella-zoster virus glycoprotein genes.

Antibodies were raised in rabbit against the in vitro translation products encoded by the varicella-zoster virus (VZV) glycoprotein genes gpI and gpIV. The antisera neutralized VZV infectivity and specifically identified two late VZV glycoproteins, gpI and gpIV, in VZV-infected cells and in the envelope of VZ virions. Pulse-chase experiments revealed a 55K precursor protein to gpIV (60K) and a 82K precursor protein to gpI (95K). Immunoprecipitation of 32P-labeled VZV-infected cells showed that the precursor-products of gpI are phosphorylated. These results demonstrate that translation products synthesized in vitro can be used to produce antibodies that recognize native viral proteins and therefore facilitate the identification and analysis of viral gene products in the infected cells.

Animals↗

PubMed1986

Polypeptides encoded by varicella-zoster virus unique short sequences.

The SalI-I and K DNA fragments which lie within the unique short sequences (Us) and contain a portion of the inverted repeat sequences (IRs/TRs) of varicella-zoster virus (VZV) DNA were cloned in an in vitro transcription vector system (pGEM-2). RNA was transcribed from both strands, translated in vitro and analyzed by SDS-PAGE. The results showed that SalI-I and K each codes for three primary translation products. Polypeptides with Mrs of 19,000 (19K), 47K, 93K/90K are encoded by SalI-I and polypeptides of 12K, 19K, and 50K are encoded by SalI-K. These results are consistent with the predicted genetic expression of the VZV SalI-I and SalI-K DNA fragments.

Cloning, Molecular↗

PubMed1986

New common nomenclature for glycoprotein genes of varicella-zoster virus and their glycosylated products.

The accumulation of recent data concerning the reactivity of monoclonal antibodies with particular varicella-zoster virus (VZV) glycoproteins and the mapping of several of their respective genes on the VZV genome has led to a unified nomenclature for the glycoprotein genes of VZV and their mature glycosylated products. Homologs to herpes simplex virus glycoprotein genes are noted.

Genes, Viral↗