The molecular genetics of herpes simplex virus latency and pathogenesis: a puzzle with many pieces still missing.
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Animals↗
Biomedical subjects
D H Gilden
Publications and source records attributed to D H Gilden.
Configuration and terminal sequences of the simian varicella virus genome.
The simian varicella virus (SVV) genome is a linear DNA molecule consisting of a unique short (Us) and a unique long (UL) region. The Us is bounded by internal (IR) and terminal inverted repeats and inverts such that virion DNA contains equimolar amounts of two genome isomers. We have sequenced the right and leftward termini and the UL-IR junction region of the SVV genome. A sequence motif common to other herpesviruses, consisting of A and T residues surrounded by G+C-rich regions, was found near the rightward terminus of the SVV genome. Sequence analysis showed no repeats surrounding the UL region of the genome. Nucleic acid hybridization and polymerase chain reaction (PCR) amplification using primers from the right and leftward ends of the SVV genome indicated that the UL region inverts. PCR amplification also showed that, compared with virion DNA, SVV genomes with connected termini are increased in infected cell DNA, suggesting the presence of circular or concatemeric genomic molecules.
Base Sequence↗
The vasculopathy of varicella-zoster virus encephalitis.
Varicella-zoster virus (VZV) encephalitis has become more prevalent in the era of acquired immunodeficiency syndrome and other immunosuppressive diseases and poses diagnostic and therapeutic challenges for clinicians, radiologists, and pathologists. Six cases studied at our institutions shed light on the patterns and pathogenesis of the disease. VZV encephalitis is predominantly a vasculopathy, involving small and large vessels, that generates seizures, mental changes, and focal deficits. Brain imaging reveals large and small ischemic or hemorrhagic infarcts, often both, of cortex and subcortical gray and white matter. Deep-seated white matter lesions often predominate and are ischemic and/or demyelinative, depending on the size of blood vessels involved and the amount of additional demyelination caused by infection of oligodendrocytes. The demyelinative lesions are smaller and less coalescent than those seen in progressive multifocal leukoencephalopathy.
Adult↗
In situ polymerase chain reaction detection of varicella zoster virus in infected cells in culture.
Polymerase chain reaction (PCR) technology provides a means of identifying genes and studying their expression with specificity and precision. Combined with in situ hybridization (ISH), the amplified gene can be localized within cells. Cell gene sequences (human alpha-tubulin) were identified by PCR-ISH first in uninfected cells in culture, then in human blood mononuclear cells, and finally in sections of human liver. Varicella zoster virus (VZV) DNA was detected for the first time in virus-infected cells in culture by PCR-ISH, and compared to ISH alone. Efficient PCR-ISH was achieved by fixation of cells or tissue sections with 2% paraformaldehyde, by amplification under conditions of complete exposure of the samples to the reagents, and by detection of amplified products using non-radioactive digoxigenin-labeled oligonucleotide probes internal to the amplified DNA segment. PCR-ISH increased the detection of VZV DNA in infected cells 5-fold compared to ISH alone. Compared to ISH alone, PCR-ISH enhances significantly the detection of virus DNA in infected cells.
Animals↗
Configuration of latent varicella-zoster virus DNA.
The configuration of latent varicella-zoster virus (VZV) DNA was analyzed by PCR. Template DNA for both internal and terminal VZV primers was present in a 1:1 ratio in ganglionic DNA, compared with a 15:1 ratio in DNA extracted from VZV virions, indicating that the VZV genomic termini are adjacent in latently infected human ganglia.
Adult↗
Varicella-zoster virus (VZV) transcription during latency in human ganglia: prevalence of VZV gene 21 transcripts in latently infected human ganglia.
Reverse transcriptase-linked PCR was used to determine the prevalence of varicella-zoster virus (VZV) gene 21 transcription in latently infected human ganglia. Under conditions wherein reverse transcriptase-linked PCR detected > or = 1,000 transcripts, VZV gene 21 RNA, but not VZV gene 40 RNA, was found in ganglia but not other tissues from five of seven humans.
Base Sequence↗
Persistence of varicella-zoster virus DNA in elderly patients with postherpetic neuralgia.
The most common complication of zoster in the elderly is postherpetic neuralgia, operationally defined as pain persisting longer than 1-2 months after rash. The cause of postherpetic neuralgia is unknown. Using polymerase chain reaction, we detected varicella zoster virus DNA in blood mononuclear cells from 11 of 51 postherpetic neuralgia patients, but not in any of 19 zoster patients without postherpetic neuralgia, or in any of 11 elderly individuals without a history of zoster. Blood mononuclear cells from nine of 27 serially-bled postherpetic neuralgia patients were positive for varicella zoster virus DNA; six were positive once, and three patients were positive more than once. Our results indicated that postherpetic neuralgia may be related to persistence of varicella zoster virus.
Aged↗
Herpes zoster with postherpetic neuralgia--persisting pain and frustration.
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Acyclovir↗
Zoster sine herpete, a clinical variant.
Two otherwise healthy immunocompetent men, ages 62 and 66 years, experienced years of radicular pain without zoster rash. An extensive search for systemic disease and malignancy was negative. Varicella-zoster virus DNA, but not herpes simplex virus DNA, was found in the cerebrospinal fluid of the first patient 5 months after the onset of pain, and in the second patient 8 months after the onset of pain. Prolonged radicular pain without zoster rash combined with the presence of varicella-zoster virus in the cerebrospinal fluid indicates that both men had zoster sine herpete, and strongly supports this syndrome as a clinical variant.
Acyclovir↗
Varicella-zoster virus (VZV) transcription during latency in human ganglia: construction of a cDNA library from latently infected human trigeminal ganglia and detection of a VZV transcript.
The entire varicella-zoster virus (VZV) genome appears to be present in latently infected human ganglia, but the extent of virus DNA transcription is unknown. Conventional methods to study virus gene transcripts by Northern (RNA) blotting are not feasible, since ganglia are small and VZV DNA is not abundant. To circumvent this problem, we prepared radiolabeled cDNA from ganglionic RNA, hybridized it to Southern blots containing VZV DNA, and demonstrated the presence of a transcript within the SalI C fragment of the virus genome (R. Cohrs, R. Mahalingam, A. N. Dueland, W. Wolf, M. Wellish, and D. H. Gilden, J. Infect. Dis. 166:S24-S29, 1992). To further map VZV transcripts, in the work described here we constructed a cDNA library from poly(A)+ RNA obtained from latently infected human ganglia. Phage DNA isolated from the library was used in PCR amplifications to detect VZV-specific inserts. The specificity of the PCRs was provided by selection of a primer specific for VZV gene 17, 18, 19, 20, or 21 and a second vector-specific primer. VZV gene 21-specific sequences were identified by PCR amplification. The PCR product contained the XhoI cloning site and poly(A)+ sequences between vector and VZV gene 21 sequences. The sequence motif at the 3' end of VZV gene 21, determined by cloning and sequencing of the PCR product, consisted of 49 to 51 nucleotide bases of 3'-untranslated DNA, the termination codon for the VZV gene 21 open reading frame, and DNA sequences reading into the VZV gene 21 open reading frame.
Aged↗
Varicella-zoster virus myelitis: an expanding spectrum.
We report four cases of varicella-zoster virus (VZV)-associated myelopathy in adults. Myelopathy was remitting-exacerbating in two remarkable instances, once acute and once chronic. VZV myelopathy was diagnosed based on the close temporal relationship between rash and onset of myelopathy, and for the first time, by polymerase chain reaction, which revealed VZV DNA in the cerebral spinal fluid of three patients with pleocytosis weeks to months later. Magnetic resonance imaging was abnormal in three of four patients. Although all four patients were treated at some time with intravenous acyclovir, concomitant treatment with steroids and the presence of acquired immunodeficiency syndrome in one patient prevented conclusions about a favorable response to therapy. Myelopathy after VZV infection may be remitting-exacerbating in addition to acute or chronic. Detection of VZV DNA in cerebral spinal fluid months after rash was useful for diagnosis and suggests a role for virus in the pathogenesis of myelopathy.
Acquired Immunodeficiency Syndrome↗
DNA sequence of a simian varicella virus gene that encodes a homologue of varicella zoster virus IE62 and herpes simplex virus ICP4.
Clinical and biological studies indicate that simian varicella virus (SVV) infection of primates is the counterpart of human varicella zoster virus (VZV) infection. We have identified an SVV open reading frame (ORF) that is homologous to the VZV protein IE62 and herpes simplex virus (HSV) ICP4. Like the genes encoding the VZV and HSV proteins, the SVV gene is located in the repeat region of the virus genome. Its expression in SVV-infected cells yields a 4.2-kb transcript. DNA sequencing of the SVV gene reveals a coding region 3834 nucleotides in length and a G+C content of 60.3%. Like ICP4 homologues in other herpesviruses, the SVV protein has five regions, two of which (regions 2 and 4) are highly conserved. The SVV protein also contains a run of serine residues in its amino terminus that is characteristic of herpesvirus ICP4 homologues. The SVV protein shows an overall homology of 24% to HSV ICP4 and 53% to VZV IE62.
Amino Acid Sequence↗
Localization of herpes simplex virus and varicella zoster virus DNA in human ganglia.
Human dorsal root ganglia from 14 randomly autopsied adults and 1 infant (all seropositive for both herpes simplex virus [HSV] and varicella zoster virus [VZV]) were examined for latent HSV-1 and VZV DNA by polymerase chain reaction. Thoracic ganglionic DNA from all subjects and trigeminal ganglionic DNA from 11 adults were analyzed. HSV-1 DNA was detected in trigeminal ganglia from 8 of 11 (73%) adults and in thoracic ganglia from 2 of 14 (14%) adults. VZV DNA was detected in trigeminal ganglia from 10 of 11 (91%) adults and in thoracic ganglia from 12 of 14 (86%) adults. None of the DNA samples were positive with primers specific for HSV-2. These findings indicate the presence of latent HSV-1 and VZV DNA in trigeminal ganglia and latent VZV DNA in thoracic ganglia of most seropositive adults. Furthermore, although HSV-1 latency most commonly develops in trigeminal ganglia, we also show for the first time the presence of HSV-1 latency in thoracic ganglia. Finally, both viruses can become latent in the same trigeminal ganglion.
Base Sequence↗
Prevalence and distribution of latent simian varicella virus DNA in monkey ganglia.
We used polymerase chain reaction to analyze the prevalence and distribution of latent simian varicella virus (SVV) in ganglionic and nonganglionic tissues from nine African green monkeys experimentally infected with SVV. Primers specific for three different regions of the SVV genome were used for amplification. SVV DNA sequences were detected in trigeminal ganglia from seven of nine monkeys and in thoracic ganglia from seven of nine monkeys. Analysis of DNA from nonneuronal tissues of three monkeys and from adrenal glands of nine monkeys revealed the presence of SVV-specific sequences in the adrenal gland of one monkey. The results indicate that, like human varicella, SVV becomes latent primarily in ganglia at multiple levels of the neuraxis, and more than one region of the SVV genome is present in latently infected ganglia. SVV latency in primates may be a useful model for varicella latency in humans.
Animals↗
Molecular analysis of simian varicella virus DNA.
Clinical and pathological studies indicate that simian varicella virus (SVV) infection in primates is the counterpart of human varicella zoster virus (VZV) infection. The SVV and VZV genomes are also similar in size and structure. To extend studies of SVV DNA, we analyzed virus DNA from African green monkey kidney cells infected with the Delta-herpes-virus strain of SVV. The infectivity of SVV DNA was 88 PFU/micrograms. The buoyant density of SVV DNA, determined by isopycnic banding in CsCl gradients, was 1.700 +/- 0.002 g/ml, corresponding to a G + C molar ratio of 40.8%. The size of SVV DNA, estimated by analysis of restriction endonuclease digestion products and pulsed-field gel electrophoresis was 125.1 and 124.9 kbp, respectively. Electron microscopy of SVV DNA revealed a long region of 110.0 kbp, a unique short (Us) region of 5.1 kbp, and inverted repeat regions of 7.5 kbp flanking the Us. An EcoRI map of SVV DNA revealed two fragments not previously reported; our complete Pstl map also shows some differences. Mapping of SVV DNA with an additional restriction enzyme, measurement of full-length SVV DNA molecules, and the first use of pulsed-field electrophoresis to size SVV DNA, confirm and extend Gray's recent finding that SVV DNA has the same size and molecular configuration as VZV. We also show for the first time that the density of SVV DNA is similar to that of VZV DNA and that SVV DNA is infectious.
Animals↗
Peripheral blood mononuclear cells of the elderly contain varicella-zoster virus DNA.
Peripheral blood mononuclear cells (PBMC) from humans of different ages were analyzed for DNA sequences specific for varicella-zoster virus (VZV) genes 29 and 62 by polymerase chain reaction (PCR). Neither VZV gene was detected in DNA from umbilical cord blood PBMC of 10 infants or from blood PBMC of two 3-year-old children. In 22 humans less than 60 years old, gene 29 was not detected, and gene 62 was detected in only one subject. In 33 humans greater than 60 years old, including patients with postherpetic neuralgia, PBMC from 4 subjects contained gene 29, 4 contained gene 62, and 1 contained both genes. The presence of VZV DNA correlated significantly with age (P less than .05, chi 2 and logistic regression analysis), but not with gender or postherpetic neuralgia.
Adult↗
Restricted transcription of varicella-zoster virus in latently infected human trigeminal and thoracic ganglia.
Normal human trigeminal and thoracic ganglia latently infected with varicella-zoster virus (VZV) were identified by polymerase chain reaction (PCR). Total RNA was extracted from these ganglia and treated with DNase until ganglionic RNA was free of VZV DNA as determined by PCR. Radiolabeled cDNA synthesized by priming with random oligonucleotides was hybridized to Southern blots containing recombinant clones that spanned greater than 95% of the VZV genome. The single region of the VZV genome detected was the 12.5-kb SalI C fragment located in the unique long segment of the viral genome. Two additional regions of the VZV genome, EcoRI G and SalI B, were detected in RNA from adult dorsal root ganglia and infant nervous system tissue.
Adult↗
Varicella-zoster virus reactivation without rash.
Reactivation of varicella-zoster virus (VZV) leads to localized zoster (shingles), a syndrome characterized by pain and a vesicular rash. Rarely, patients experience radicular pain without zosteriform rash, cases that have been regarded as zoster sine herpete (zoster without rash). Virologic evidence for zoster sine herpete is sparse. However, VZV can produce other neurologic and visceral diseases in the absence of rash or radicular pain. The clinical and virologic features of zoster sine herpete and other disorders produced by VZV without rash are reviewed. Evidence is also presented for the detection of VZV DNA in human blood mononuclear cells of elderly individuals in the absence of skin lesions or other VZV-associated neurologic or systemic disease.
Aged↗