Previous Article | Next Article 
Journal of Virology, December 1999, p. 10514-10518, Vol. 73, No. 12
0022-538X/99/$04.00+0
Quantitation of Latent Varicella-Zoster Virus and
Herpes Simplex Virus Genomes in Human Trigeminal Ganglia
Stephanie R.
Pevenstein,1
Richard K.
Williams,1
Daniel
McChesney,1
Erik K.
Mont,2
John E.
Smialek,3 and
Stephen
E.
Straus1,*
Medical Virology Section, Laboratory of
Clinical Investigation, National Institute of Allergy and Infectious
Diseases,1 and Anatomical Pathology
Service, Laboratory of Pathology, National Cancer
Institute,2 National Institutes of Health,
Bethesda, and Department of Pathology, University of Maryland
School of Medicine, Baltimore,3 Maryland
Received 10 May 1999/Accepted 31 August 1999
 |
ABSTRACT |
Using real-time fluorescence PCR, we quantitated the numbers of
copies of latent varicella-zoster virus (VZV) and herpes simplex virus
type 1 (HSV-1) and type 2 (HSV-2) genomes in 15 human trigeminal ganglia. Eight (53%) and 1 (7%) of 15 ganglia were PCR positive for
HSV-1 or -2 glycoprotein G genes, with means of 2,902 ± 1,082 (standard error of the mean) or 109 genomes/105 cells,
respectively. Eleven of 14 (79%) to 13 of 15 (87%) of the ganglia
were PCR positive for VZV gene 29, 31, or 62. Pooling of the results
for the three VZV genes yielded a mean of 258 ± 38 genomes/105 ganglion cells. These levels of latent viral
genome loads have implications for virus distribution in and
reactivation from human sensory ganglia.
| |
TEXT |
Herpes simplex virus types 1 and 2 (HSV-1 and -2) and varicella-zoster virus (VZV) are alphaherpesviruses
that infect, establish latency in, and subsequently reactivate from
human sensory nerve ganglia (1, 36). Following reactivation
from latent ganglia reservoirs, each of these herpesviruses may cause
significant clinical disease in the individual and may spread to
uninfected persons. Symptomatic VZV reactivation is an infrequent,
usually once-in-a-lifetime event that results in zoster (shingles),
while HSV-1 and -2 reactivation occurs frequently and results in
numerous symptomatic and asymptomatic recurrences of oral and genital herpes.
Little is known regarding the mechanism underlying the particular
patterns of latency and reactivation that distinguish HSV-1 and -2 infection from that with VZV. Abundant data from both human studies and
animal models confirm that HSV-1 and -2 persist in sensory neurons but
that satellite glial cells are spared from harboring latent HSV
(7, 29, 30). Data regarding the site of VZV latency have
been controversial, with various reports indicating it to be neurons,
nonneuronal cells, or both (7, 9, 17, 22, 26). Moreover,
estimates of the proportions of cells harboring HSV and VZV and the
quantity of latent viral DNA in ganglia have varied widely (7, 17,
22, 25, 30). Recent animal studies show that latent viral genome
levels in sensory ganglia influence the reactivation frequency of HSV-1
and -2, suggesting that the quantity of latent viral genome copies per
ganglion
the latent viral loadmay be a significant determinant of
herpesvirus reactivation from the nervous system (21, 31,
32). To further clarify the nature and distribution of latent HSV
and VZV genomes in human ganglia, we developed and used several
sensitive and specific PCR assays.
Human tissue samples.
Human trigeminal ganglia were harvested
within 24 h postmortem, frozen in dry ice, and stored at 
70°C
until DNA extraction. The general clinical histories and causes of
death are summarized in Table 1.
DNA was extracted by the protocol described in the instructions for a
Puregene DNA isolation kit (D-5000A; Gentra Systems, Minneapolis,
Minn.) with a few modifications. Ganglia were pulverized to a fine
powder on dry ice and incubated in cell lysis buffer and 10 mg of
proteinase K per ml for 3 days to ensure complete homogenization.
Following protein precipitation, DNA was ethanol precipitated and
resuspended in water. DNA concentration was estimated by
spectrophotometry, and purity was determined from the ratios of the
optical density at 260 nm to that at 280 nm. On average, the ganglia
yielded 478 ± 46 (mean ± standard error of the mean [SEM]) µg of DNA.
QF-PCR assays.
Quantitative fluorescent (QF) PCR was performed
with a Prism 7700 sequence detector (PE Applied Biosystems, Foster
City, Calif.) according to supplied guidelines for real-time DNA
amplification. Real-time PCR relies on a quantitative increase in
fluorescence due to cleavage of a 5' reporter dye from a dually labeled
fluorogenic probe oligonucleotide by the 5'
3' nuclease activity of
Taq DNA polymerase.
The genes encoding VZV glycoprotein B (gB; open reading frame [ORF]
31), ORF 62 (which encodes the major immediate early transactivator),
ORF 29 (which encodes a putative early major DNA-binding protein),
HSV-1 glycoprotein G (gG1), and HSV-2 glycoprotein G (gG2) were
selected for quantification. The forward and reverse primers and
probe
for the VZV gB gene were described by Kimura et al. (
18)
(Table
2). The forward and reverse
primers and probes for VZV
ORF 29 and ORF 62 and for the gG genes of
HSV-1 and HSV-2 were
designed with the Primer Express program (PE
Applied Biosystems)
(Table
2) and synthesized by Bioserve
Biotechnologies (Laurel,
Md.). QF PCR was also performed with the
primers and the probe
(Table
2) provided with the Taqman
-actin
reagent kit (PE Applied
Biosystems) to normalize each of the ganglion
extracts for amplifiable
human DNA.
All PCRs were performed in triplicate on two separate occasions with a
Taqman PCR kit (PE Applied Biosystems) in the absence
of reverse
transcriptase, so that only DNA was amplified. Each
50-µl PCR mixture
contained 500 ng of human trigeminal ganglion
DNA with final
concentrations of each primer of 1,000 nM and of
each probe sequence of
200 nM. Multiple human trigeminal DNA samples
were run together on a
plate in parallel with duplicate sets of
DNA standards. Each primer set
mixture was run on a separate plate.
Standard curves for each targeted
viral gene were generated by
mixing serial 10-fold dilutions of
plasmids containing 10
0 to 10
6 copies of the
desired genes. Additional dilutions within that
range were analyzed for
plasmids bearing the genes for gB, ORF
62, and gG1 to better delineate
the sensitivity limits at the
low ends of the standard curves.
Uninfected BALB/c mouse trigeminal
DNA (500 ng) was added to all
plasmid standards to compensate
for the potential inhibition of
amplification reactions by added
DNA. For VZV gB, the gB gene contained
in the pUC19 vector, a
gift of Liyanage Perera, was used. For ORF 29 and ORF 62, their
respective
EcoRI-B and
EcoRI-A
fragments from the VZV strain Ellen
contained in the pGEM vector were
used. Plasmids containing the
HSV-1 and HSV-2 gG genes were obtained
from Mark Challberg and
Philip Krause, respectively. In addition, no
template control
reaction mixtures containing the appropriate probe and
primer
system without DNA were run on all plates in triplicate. PCR
mixtures
were subjected to 2 min at 50°C (reaction of AmpErase
uracil-
N-glycosylase),
10 min at 95°C (activation of
AmpliTaq Gold), and 55 cycles of
15 s at 95°C and 1 min at
60°C. For
-actin amplification, 40
cycles were
performed.
For each reaction, real-time fluorescence values were measured as a
function of the quantity of a reporter dye (6-carboxy-fluorescein
[FAM]) released during amplification. A threshold cycle
(
Ct) value
for each sample was determined as the
number of the first cycle
at which the measured fluorescence exceeded
the threshold limit
(10 times the standard deviation of the baseline).
Ct values observed
for human trigeminal DNA
samples were used to calculate the viral
genome copy number for each
sequence amplified based on the standard
curves for plasmids containing
test
sequence.
Standard curves for three VZV-containing plasmids are shown in Fig.
1 as examples. No cross-reactivity was
observed between
any of the viral DNA assays. The limits of sensitivity
of the
QF PCRs for all of the viral genes were estimated as the lowest
plasmid dilutions that yielded comparable
Ct
values in replicate
samples: they were all <10 copies/500 ng of input
DNA. The human
-actin standards were not evaluated below 100 copies/500 ng of
DNA. All standard curves were fit by linear
regression, with correlation
coefficients of >0.92.
View larger version (19K):
[in this window]
[in a new window]
|
FIG. 1.
Standard curves for QF-PCR assays for VZV genes encoding
ORFs 29 and 62 and gB. Shown are the Ct values
at which a given input of a VZV DNA-containing plasmid was detected by
PCR. Plasmid concentrations tested ranged from 100 to
106 copies per reaction mixture. The Spearman rank
coefficient of correlation (r) for lines fitting each
standard curve are given.
|
|
Ct values for plasmid dilutions in these
standard curves were compared with
Ct values of
human trigeminal ganglia samples
to estimate the number of copies of
each viral gene present in
the approximately 500 ng of extracted DNA
analyzed for each reaction.
To more accurately estimate the amount of
human genomic DNA in
each trigeminal ganglion extract, we quantitated
the number of
copies of
-actin genes in that sample volume and
normalized the
number of DNA copies observed to the number of copies
per 2 ×
10
5 -actin copies, i.e., per
10
5 cells. Because the mean number of copies of
-actin
quantified
in 500 ng of DNA was 6.4 × 10
4 ± 0.8 × 10
4, we calculated that each cell contained
15.6 pg of
DNA.
VZV and HSV latent DNA load.
The numbers of copies of the
various VZV and HSV genes per 105 cells are displayed in
Fig. 2 along with tabulations of their mean numbers (± SEM) for ganglia that yielded positive results (
10
copies/500 ng of DNA). By these criteria, the proportions (and
percentages) of ganglia positive for each viral gene were as follows:
11 of 14 were positive (79%) for the ORF 29 gene, 12 of 15 were
positive (80%) for the gB gene, 13 of 15 were positive (87%) for the
ORF 62 gene, 8 of 15 were positive (53%) for the gG1 gene, and 1 of 14 were (7%) positive for the gG2 gene.
View larger version (18K):
[in this window]
[in a new window]
|
FIG. 2.
Copies of VZV, HSV-1, and HSV-2 genes in human
trigeminal ganglia. The mean ± SEM number of copies per
105 cells for each gene in the PCR-positive ganglia are
tabulated below the figure. The filled circles show the gene copy
numbers for samples that exceeded the assay limit of detection (10
copies/500 ng of total DNA). Open circles represent values for samples
for which gene copy numbers may have been extrapolated from the QF-PCR
results but were below the threshold of reliable DNA detection (<10
copies/500 ng of DNA). The inability to define a precise threshold for
positive ganglia on the per-105-cell basis used here
reflects the variability in -actin gene number per 500 ng of total
DNA from the individual ganglia.
|
|
By the Spearman rank test, the numbers of copies of all three VZV genes
correlated highly significantly with each other (
P < 0.001). There were no statistical associations between HSV gene
copy numbers and VZV gene numbers. Moreover, the mean number of
VZV ORF
62 genes, 2.7 ± 0.4 times the numbers of the gB and ORF
29 genes,
was not dissimilar from the expected ratio of 2.0 for
this diploid
viral gene. With the numbers obtained for gB and
ORF 29 and one-half of
the number obtained for the diploid ORF
62 gene being pooled, the
QF-PCR assay revealed that PCR-positive
ganglia contained a mean of
258 ± 38 VZV genome copies/10
5 cells. This value was
significantly less than the 2,902 ± 1,082
HSV-1 genomes per
10
5 cells in positive ganglia (
P = 0.02;
Wilcoxon rank sum test)
but comparable to the copy number of genomes in
the one ganglion
containing detectable HSV-2 DNA, 109 genomes/10
5 cells. Latent viral DNA loads were not grossly
different for
ganglia obtained from individuals known to be
immunocompromised
and those presumed to be
immunocompetent.
Implications of the data.
The detection of VZV DNA in 79 to
87% of ganglia is in concordance with the very high proportion of
American adults who are seropositive for VZV and with data obtained
from prior standard nonquantitative PCR assays and in situ
hybridization (1, 13, 27). Slightly more than half of the
ganglia were HSV-1 DNA positive, also commensurate with the
seroprevalence of 50 to 70% reported for this virus for young American
adults and with prior molecular studies of human ganglia (7, 11,
36). HSV-2 DNA was detected in only 1 of 15 ganglia, reflecting
the relatively low rate of facial infection with this virus
(36).
Extrapolations of our data permit estimates of the total copy numbers
and distributions of these three viruses in human trigeminal
ganglia.
We recovered a mean of 478 µg of DNA per ganglion. Based
on our
estimate of 15.6 pg of DNA/cell, the average trigeminal
ganglion
contained upward of 3 × 10
7 cells. Thus, we estimate
that, on average, each ganglion latently
infected with VSV, HSV-1, or
HSV-2 contained 7.7 × 10
4, 8.7 × 10
5, or 3.2 × 10
4 genomes, respectively.
Our estimate for latent HSV-1 load in
human ganglia (Fig.
2) is similar
to that reported by Efstathiou
et al. (
11) (1,000 to 10,000 copies), but the estimate for VZV
load is higher than the 6 to 31 copies/10
5 cells reported by Mahalingam et al. using the
less precise PCR
and Southern hybridization methods (
26).
Ball et al. reported that human trigeminal ganglia contain an average
of 8.1 × 10
4 neurons (
2). Since HSV-1 and
-2 persist exclusively in neurons
(
5,
7,
8,
29,
30,
34,
35),
we can project that
each latently infected neuron contains at least 11 copies of HSV-1
DNA, assuming all neurons are infected. While in situ
hybridization
studies for HSV-1 latency-associated transcripts
suggested that
only 1 to 4% of neurons are positive (
7,
8,
10,
34,
35), the more recent PCR and in situ-PCR analyses
demonstrated
that 3 to 10 times that percentage are positive
(
6-8,
29-32,
34,
35). These data indicate that latently
infected trigeminal neurons
contain an average of 28 or more HSV-1
genomes, a copy number
similar to those of Epstein-Barr virus episomes
carried in B cell
lines (
12,
28) and papillomavirus episomes
in human cervical
cancer cells (
3,
4,
15,
16).
The low copy number of detectable HSV-2 DNA in trigeminal ganglia may
parallel the very low frequency with which this virus
recurs following
initial infection in the mouth (
20). Since
the latent viral
load correlates well with recurrence rates in
infected animals, if
further studies confirm the low HSV-2 copy
number in human ganglia,
these data suggest that the same is also
true for humans (
21,
24,
31,
32). If so, some obstacle
must impede infection and
establishment of latency of HSV-2 in
human trigeminal
ganglia.
Although our initial data suggested that VZV persists only in
nonneuronal cells (
7), recent data compel us to conclude
that both neurons and nonneuronal cells are infected by VZV both
during
productive infection (
7,
22) and during latency (
7,
9,
14,
17,
22,
25). Were VZV to persist exclusively
in neurons, the
average of 7.7 × 10
4 genome copies we detected could
be distributed among nearly all
of the estimated 8.1 × 10
4 neurons; however, they are not distributed in this way
(
17,
22). While we found no published estimates of the
numbers of
satellite and other nonneuronal cell populations in human
ganglia,
sensory neurons are very large (50 to 100 µm in diameter)
and
are encircled and contacted by about 10 nucleated satellite cells
each in most thin (6 to 8-µm) histologic sections (unpublished
data
and references
6,
10, and
19),
which implies that
there are at least 100 satellite cells surrounding
every neuron
or at least 8 × 10
6 satellite cells per
human trigeminal
ganglion.
If we were to assume that VZV persisted in the same proportions of
neurons and nonneuronal cells and at roughly the same copy
number per
cell as that of HSV-1 (
28), fewer than 1 in 1,000
total cells would
prove positive. Lungu et al. (
22,
23) estimated
that 5 to
30% of both neurons and satellite cells are positive
for VZV DNA or
VZV proteins, an estimate that is far higher than
that permitted by
these data. Kennedy et al. (
17), however,
estimated by in
situ PCR that 2% of neurons (~1,600 cells according
to the estimate
of Ball et al. [
2]) and 0.1% of satellite cells
(~8,000 cells according to our estimate) are VZV DNA positive.
If the
estimate of Kennedy et al. is correct, the 7.7 × 10
4
VZV genomes could persist in this number of cells at a density
of eight
copies/cell, not dissimilar from that for HSV-1. We are
currently
testing the validity of these extrapolations from our
QF-PCR data by
analyzing sections microdissected from human ganglia
(
33).
The present data provide refined estimates of the proportion of human
trigeminal ganglia containing VZV, HSV-1, and HSV-2
DNAs and the latent
DNA loads for each of these neurotropic viruses.
Moreover, they have
implications regarding the distribution of
these three viruses in human
ganglia and the pathogenesis of reactivation
infections associated with
them.
| |
ACKNOWLEDGMENTS |
We thank Jeffrey Cohen, Philip Krause, and Nancy Tresser for advice
and assistance in this project, Peter Kennedy for additional comments
on the manuscript, and Uri Lopatin for help with statistical analyses.
| |
FOOTNOTES |
*
Corresponding author. Mailing address: LCI, NIAID, NIH,
Building 10, 11N228, 10 Center Dr., Bethesda, MD 20892. Phone: (301) 496-5807. Fax: (301) 496-7383. E-mail: sstraus{at}nih.gov.
| |
REFERENCES |
| 1.
|
Arvin, A. M.
1996.
Varicella-zoster virus, p. 2547-2585.
In
B. N. Fields, D. M. Knipe, and P. M. Howley (ed.), Fields virology, 3rd ed., vol. 2. Lippincott-Raven, Philadelphia, Pa
|
| 2.
|
Ball, M. J.,
K. Nuttall, and K. G. Warren.
1982.
Neuronal and lymphocytic populations in human trigeminal ganglia: implications for aging and for latent virus.
Neuropathol. Appl. Neurobiol.
8:177-187[Medline].
|
| 3.
|
Berumen, J.,
L. Casas,
E. Segura,
J. L. Amezcua, and A. Garcia-Carranca.
1994.
Genome amplification of human papillomavirus types 16 and 18 in cervical carcinomas is related to the retention of E1/E2 genes.
Int. J. Cancer
56:640-645[Medline].
|
| 4.
|
Brown, D. R.,
J. T. Bryan,
H. Cramer,
B. P. Katz,
V. Handy, and K. H. Fife.
1994.
Detection of multiple human papillomavirus types in condylomata acuminata from immunosuppressed patients.
J. Infect. Dis.
170:759-765[Medline].
|
| 5.
|
Croen, K. D.,
J. M. Ostrove,
L. Dragovic, and S. E. Straus.
1991.
Characterization of herpes simplex virus type 2 latency-associated transcription in human sacral ganglia and in cell culture.
J. Infect. Dis.
163:23-28[Medline].
|
| 6.
|
Croen, K. D.,
J. M. Ostrove,
L. J. Dragovic,
J. E. Smialek, and S. E. Straus.
1987.
Latent herpes simplex virus in human trigeminal ganglia. Detection of an immediate early gene "anti-sense" transcript by in situ hybridization.
N. Engl. J. Med.
317:1427-1432[Abstract].
|
| 7.
|
Croen, K. D.,
J. M. Ostrove,
L. J. Dragovic, and S. E. Straus.
1988.
Patterns of gene expression and sites of latency in human nerve ganglia are different for varicella-zoster and herpes simplex viruses.
Proc. Natl. Acad. Sci. USA
85:9773-9777[Abstract/Free Full Text].
|
| 8.
|
Deatly, A., and A. Lumsden.
1984.
RNA from an immediate early region of the type 1 herpes simplex virus genome is present in the trigeminal ganglia of latently infected mice.
Proc. Natl. Acad. Sci. USA
84:3204-3208.
|
| 9.
|
Dueland, A. N.,
T. Ranneberg-Nilsen, and M. Degre.
1995.
Detection of latent varicella zoster virus DNA and human gene sequences in human trigeminal ganglia by in situ amplification combined with in situ hybridization.
Arch. Virol.
140:2055-2066[Medline].
|
| 10.
|
Ecob-Prince, M. S.,
C. M. Preston,
F. J. Rixon,
K. Hassan, and P. G. E. Kennedy.
1993.
Neurons containing latency-associated transcripts are numerous and widespread in dorsal root ganglia following footpad inoculation of mice with herpes simplex virus type 1 mutant in 1814.
J. Gen. Virol.
74:985-994[Abstract/Free Full Text].
|
| 11.
|
Efstathiou, S.,
A. C. Minson,
H. J. Field,
J. R. Anderson, and P. Wildy.
1986.
Detection of herpes simplex virus-specific DNA sequences in latently infected mice and in humans.
J. Virol.
57:446-455[Abstract/Free Full Text].
|
| 12.
|
Ernberg, I.,
M. Andersson-Anvret, and G. Klein.
1977.
Relationship between amount of Epstein-Barr virus-determined nuclear antigen per cell and number of EBV-DNA copies per cell.
Nature
226:269-271.
|
| 13.
|
Furuta, Y.,
T. Takasu,
S. Fukuda,
K. C. Sato-Matsumura,
Y. Inuyama,
R. Hondo, and K. Nagashima.
1992.
Detection of varicella-zoster virus DNA in human geniculate ganglia by polymerase chain reaction.
J. Infect. Dis.
166:1157-1159[Medline].
|
| 14.
|
Gilden, D. H.,
A. Vafai,
Y. Shtram,
Y. Becker,
M. Devlin, and M. Wellish.
1983.
Varicella-zoster virus DNA in human sensory ganglia.
Nature
306:478-480[Medline].
|
| 15.
|
Ikenberg, H.,
M. Runge,
A. Goppinger, and A. Pfleiderer.
1990.
Human papillomavirus DNA in invasive carcinoma of the vagina.
Obstet. Gynecol.
76:432-438[Medline].
|
| 16.
|
Jeon, S.,
B. L. Allen-Hoffmann, and P. F. Lambert.
1995.
Integration of human papillomavirus type 16 into the human genome correlates with a selective growth advantage of cells.
J. Virol.
69:2989-2997[Abstract].
|
| 17.
|
Kennedy, P. G.,
E. Grinfeld, and J. W. Gow.
1998.
Latent varicella-zoster virus is located predominantly in neurons in human trigeminal ganglia.
Proc. Natl. Acad. Sci. USA
95:4658-4662[Abstract/Free Full Text].
|
| 18.
|
Kimura, H.,
Y. Wang,
L. Pesnicak,
J. I. Cohen,
J. J. Hooks,
S. E. Straus, and R. K. Williams.
1998.
Recombinant varicella-zoster virus glycoproteins E and I: immunologic responses and clearance of virus in a guinea pig model of chronic uveitis.
J. Infect. Dis.
178:310-317[Medline].
|
| 19.
|
Krause, P. R.,
K. D. Croen,
S. E. Straus, and J. M. Ostrove.
1988.
Detection and preliminary characterization of herpes simplex virus type 1 transcripts in latently infected human trigeminal ganglia.
J. Virol.
62:4819-4823[Abstract/Free Full Text].
|
| 20.
|
Lafferty, W. E.,
R. W. Coombs,
J. Benedetti,
C. Critchlow, and L. Corey.
1987.
Recurrences after oral and genital herpes simplex virus infection. Influence of site of infection and viral type.
N. Engl. J. Med.
316:1444-1449[Abstract].
|
| 21.
|
Lekstrom-Himes, J. A.,
L. Pesnicak, and S. E. Straus.
1998.
The quantity of latent viral DNA correlates with the relative rates at which herpes simplex virus types 1 and 2 cause recurrent genital herpes outbreaks.
J. Virol.
72:2760-2764[Abstract/Free Full Text].
|
| 22.
|
Lungu, O.,
P. W. Annunziato,
A. Gershon,
S. M. Staugaitis,
D. Josefson,
P. LaRussa, and S. J. Silverstein.
1995.
Reactivated and latent varicella-zoster virus in human dorsal root ganglia.
Proc. Natl. Acad. Sci. USA
92:10980-10984[Abstract/Free Full Text].
|
| 23.
|
Lungu, O.,
C. A. Panagiotidis,
P. W. Annunziato,
A. A. Gershon, and S. J. Silverstein.
1998.
Aberrant intracellular localization of varicella-zoster virus regulatory proteins during latency.
Proc. Natl. Acad. Sci. USA
95:7080-7085[Abstract/Free Full Text].
|
| 24.
|
Maggioncalda, J.,
A. Mehta,
Y. H. Su,
N. W. Fraser, and T. M. Block.
1996.
Correlation between herpes simplex virus type 1 rate of reactivation from latent infection and the number of infected neurons in trigeminal ganglia.
Virology
225:72-81[Medline].
|
| 25.
|
Mahalingam, R.,
M. Wellish,
R. Cohrs,
S. Debrus,
J. Piette,
B. Rentier, and D. Gilden.
1996.
Expression of protein encoded by varicella-zoster virus open reading frame 63 in latently infected human ganglionic neurons.
Proc. Natl. Acad. Sci. USA
93:2122-2124[Abstract/Free Full Text].
|
| 26.
|
Mahalingam, R.,
M. Wellish,
D. Lederer,
B. Forghani,
R. Cohrs, and D. Gilden.
1993.
Quantitation of latent varicella-zoster virus DNA in human trigeminal ganglia by polymerase chain reaction.
J. Virol.
67:2381-2384[Abstract/Free Full Text].
|
| 27.
|
Mahalingam, R.,
M. C. Wellish,
A. N. Dueland,
R. J. Cohrs, and D. H. Gilden.
1992.
Localization of herpes simplex virus and varicella zoster virus DNA in human ganglia.
Ann. Neurol.
31:444-448[Medline].
|
| 28.
|
Nonoyama, M., and J. S. Pagano.
1973.
Homology between Epstein-Barr virus DNA and viral DNA from Burkitt's lymphoma and nasopharyngeal carcinoma determined by DNA-DNA reassociation kinetics.
Nature
242:44-47[Medline].
|
| 29.
|
Ramakrishnan, R.,
P. L. Poliani,
M. Levine,
J. C. Glorioso, and D. J. Fink.
1996.
Detection of herpes simplex virus type 1 latency-associated transcript expression in trigeminal ganglia by in situ reverse transcriptase PCR.
J. Virol.
70:6519-6523[Abstract].
|
| 30.
|
Sawtell, N. M.
1997.
Comprehensive quantification of herpes simplex virus latency at the single-cell level.
J. Virol.
71:5423-5431[Abstract].
|
| 31.
|
Sawtell, N. M.
1998.
The probability of in vivo reactivation of herpes simplex virus type 1 increases with the number of latently infected neurons in the ganglia.
J. Virol.
72:6888-6892[Abstract/Free Full Text].
|
| 32.
|
Sawtell, N. M.,
D. K. Poon,
C. S. Tansky, and R. L. Thompson.
1998.
The latent herpes simplex virus type 1 genome copy number in individual neurons is virus strain specific and correlates with reactivation.
J. Virol.
72:5343-5350[Abstract/Free Full Text].
|
| 33.
|
Simone, N. L.,
R. F. Bonner,
J. W. Gillespie,
M. R. Emmert-Buck, and L. A. Liotta.
1998.
Laser-capture microdissection: opening the microscopic frontier to molecular analysis.
Trends Genet.
14:272-276[Medline].
|
| 34.
|
Stevens, J. G.,
E. K. Wagner,
G. B. Devi-Rao,
M. L. Cook, and L. T. Feldman.
1987.
RNA complementary to a herpesvirus  gene mRNA is prominent in latently infected neurons.
Science
235:1056-1059[Abstract/Free Full Text].
|
| 35.
|
Tenser, R. B.,
M. Dawson,
S. J. Ressel, and M. E. Dunstan.
1982.
Detection of herpes simplex virus mRNA in latently infected trigeminal ganglion neurons by in situ hybridization.
Ann. Neurol.
11:285-291[Medline].
|
| 36.
|
Whitley, R. J.
1996.
Herpes simplex viruses, p. 2297-2342.
In
B. N. Fields, D. M. Knipe, and P. M. Howley (ed.), Fields virology, 3rd ed., vol. 2. Lippincott-Raven, Philadelphia, Pa
|
Journal of Virology, December 1999, p. 10514-10518, Vol. 73, No. 12
0022-538X/99/$04.00+0
This article has been cited by other articles:
-
Boving, M. K., Pedersen, L. N., Moller, J. K.
(2009). Eight-Plex PCR and Liquid-Array Detection of Bacterial and Viral Pathogens in Cerebrospinal Fluid from Patients with Suspected Meningitis. J. Clin. Microbiol.
47: 908-913
[Abstract]
[Full Text]
-
Hill, J. M., Ball, M. J., Neumann, D. M., Azcuy, A. M., Bhattacharjee, P. S., Bouhanik, S., Clement, C., Lukiw, W. J., Foster, T. P., Kumar, M., Kaufman, H. E., Thompson, H. W.
(2008). The High Prevalence of Herpes Simplex Virus Type 1 DNA in Human Trigeminal Ganglia Is Not a Function of Age or Gender. J. Virol.
82: 8230-8234
[Abstract]
[Full Text]
-
Tang, S., Bertke, A. S., Patel, A., Wang, K., Cohen, J. I., Krause, P. R.
(2008). An acutely and latently expressed herpes simplex virus 2 viral microRNA inhibits expression of ICP34.5, a viral neurovirulence factor. Proc. Natl. Acad. Sci. USA
105: 10931-10936
[Abstract]
[Full Text]
-
Reichelt, M., Zerboni, L., Arvin, A. M.
(2008). Mechanisms of Varicella-Zoster Virus Neuropathogenesis in Human Dorsal Root Ganglia. J. Virol.
82: 3971-3983
[Abstract]
[Full Text]
-
Zerboni, L., Reichelt, M., Jones, C. D., Zehnder, J. L., Ito, H., Arvin, A. M.
(2007). From the Cover: Aberrant infection and persistence of varicella-zoster virus in human dorsal root ganglia in vivo in the absence of glycoprotein I. Proc. Natl. Acad. Sci. USA
104: 14086-14091
[Abstract]
[Full Text]
-
Miller, C. S., Berger, J. R., Mootoor, Y., Avdiushko, S. A., Zhu, H., Kryscio, R. J.
(2006). High prevalence of multiple human herpesviruses in saliva from human immunodeficiency virus-infected persons in the era of highly active antiretroviral therapy.. J. Clin. Microbiol.
44: 2409-2415
[Abstract]
[Full Text]
-
Hoover, S. E., Cohrs, R. J., Rangel, Z. G., Gilden, D. H., Munson, P., Cohen, J. I.
(2006). Downregulation of Varicella-Zoster Virus (VZV) Immediate-Early ORF62 Transcription by VZV ORF63 Correlates with Virus Replication In Vitro and with Latency.. J. Virol.
80: 3459-3468
[Abstract]
[Full Text]
-
Wang, K., Lau, T. Y., Morales, M., Mont, E. K., Straus, S. E.
(2005). Laser-Capture Microdissection: Refining Estimates of the Quantity and Distribution of Latent Herpes Simplex Virus 1 and Varicella-Zoster Virus DNA in Human Trigeminal Ganglia at the Single-Cell Level. J. Virol.
79: 14079-14087
[Abstract]
[Full Text]
-
Zerboni, L., Ku, C.-C., Jones, C. D., Zehnder, J. L., Arvin, A. M.
(2005). Varicella-zoster virus infection of human dorsal root ganglia in vivo. Proc. Natl. Acad. Sci. USA
102: 6490-6495
[Abstract]
[Full Text]
-
Miller, C. S., Avdiushko, S. A., Kryscio, R. J., Danaher, R. J., Jacob, R. J.
(2005). Effect of Prophylactic Valacyclovir on the Presence of Human Herpesvirus DNA in Saliva of Healthy Individuals after Dental Treatment. J. Clin. Microbiol.
43: 2173-2180
[Abstract]
[Full Text]
-
Enomoto, Y., Yoshikawa, T., Ihira, M., Akimoto, S., Miyake, F., Usui, C., Suga, S., Suzuki, K., Kawana, T., Nishiyama, Y., Asano, Y.
(2005). Rapid Diagnosis of Herpes Simplex Virus Infection by a Loop-Mediated Isothermal Amplification Method. J. Clin. Microbiol.
43: 951-955
[Abstract]
[Full Text]
-
Baiker, A., Fabel, K., Cozzio, A., Zerboni, L., Fabel, K., Sommer, M., Uchida, N., He, D., Weissman, I., Arvin, A. M.
(2004). Varicella-zoster virus infection of human neural cells in vivo. Proc. Natl. Acad. Sci. USA
101: 10792-10797
[Abstract]
[Full Text]
-
Lundberg, P., Welander, P., Openshaw, H., Nalbandian, C., Edwards, C., Moldawer, L., Cantin, E.
(2003). A Locus on Mouse Chromosome 6 That Determines Resistance to Herpes Simplex Virus Also Influences Reactivation, While an Unlinked Locus Augments Resistance of Female Mice. J. Virol.
77: 11661-11673
[Abstract]
[Full Text]
-
Levin, M. J., Cai, G.-Y., Manchak, M. D., Pizer, L. I.
(2003). Varicella-Zoster Virus DNA in Cells Isolated from Human Trigeminal Ganglia. J. Virol.
77: 6979-6987
[Abstract]
[Full Text]
-
Kaiser, P., Underwood, G., Davison, F.
(2002). Differential Cytokine Responses following Marek's Disease Virus Infection of Chickens Differing in Resistance to Marek's Disease. J. Virol.
77: 762-768
[Abstract]
[Full Text]
-
Hata, A., Asanuma, H., Rinki, M., Sharp, M., Wong, R. M., Blume, K., Arvin, A. M.
(2002). Use of an Inactivated Varicella Vaccine in Recipients of Hematopoietic-Cell Transplants. NEJM
347: 26-34
[Abstract]
[Full Text]
-
Bustos, D. E., Atherton, S. S.
(2002). Detection of Herpes Simplex Virus Type 1 in Human Ciliary Ganglia. IOVS
43: 2244-2249
[Abstract]
[Full Text]
-
Cohrs, R. J., Wischer, J., Essman, C., Gilden, D. H.
(2002). Characterization of Varicella-Zoster Virus Gene 21 and 29 Proteins in Infected Cells. J. Virol.
76: 7228-7238
[Abstract]
[Full Text]
-
Mackay, I. M., Arden, K. E., Nitsche, A.
(2002). Real-time PCR in virology. Nucleic Acids Res
30: 1292-1305
[Abstract]
[Full Text]
-
Verstrepen, W. A., Kuhn, S., Kockx, M. M., Van De Vyvere, M. E., Mertens, A. H.
(2001). Rapid Detection of Enterovirus RNA in Cerebrospinal Fluid Specimens with a Novel Single-Tube Real-Time Reverse Transcription-PCR Assay. J. Clin. Microbiol.
39: 4093-4096
[Abstract]
[Full Text]
-
Cohrs, R. J., Randall, J., Smith, J., Gilden, D. H., Dabrowski, C., van der Keyl, H., Tal-Singer, R.
(2000). Analysis of Individual Human Trigeminal Ganglia for Latent Herpes Simplex Virus Type 1 and Varicella-Zoster Virus Nucleic Acids Using Real-Time PCR. J. Virol.
74: 11464-11471
[Abstract]
[Full Text]
-
Kimura, H., Kido, S., Ozaki, T., Tanaka, N., Ito, Y., Williams, R. K., Morishima, T.
(2000). Comparison of Quantitations of Viral Load in Varicella and Zoster. J. Clin. Microbiol.
38: 2447-2449
[Abstract]
[Full Text]
Top
Abstract
Text
