Abstract
In this investigation we determined the dynamics of herpes simplex virus type 1 (HSV-1) DNA and latency-associated transcripts (LAT) in the latently infected rabbit trigeminal ganglion. Rabbit eyes were infected with either the McKrae strain or the l7Syn+ strain of HSV-1. Rabbits were sacrificed between 5 and 360 days after infection and their trigeminal ganglia were analyzed for the number of HSV DNA genomes and the number of neuronal cells expressing LAT. There was no statistically significant change in the number of HSV genomes or the number of neuronal cells expressing LAT in these ganglia between 20 and 360 days after infection. For both strains, the amount of HSV DNA averaged 16.8 genomes per 100 cells, and 9.2% of the neurons expressed LAT. There were 17 to 34 HSV genomes per LAT-expressing neuronal cell. The number of LAT-expressing neurons did not change over the 360 days. Spontaneous reactivation (HSV-1 recovery in tear film) and recurrence (HSV-1-specific epithelial lesions) occurred during the period of this study; however, these events did not alter the quantity of HSV-1 DNA or the number of LAT-expressing cells. These results suggest that after the latent infection is established, the viral DNA in the ganglia does not replicate to any measurable extent over long periods of latency, since no significant change in the number of HSV genomes occurs. The results also suggest that only a very small number of latently infected neuronal cells are needed to produce infectious HSV-1 during reactivation.
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- Berman E. J., Hill J. M. Spontaneous ocular shedding of HSV-1 in latently infected rabbits. Invest Ophthalmol Vis Sci. 1985 Apr;26(4):587–590. [PubMed] [Google Scholar]
- Beyer C. F., Hill J. M., Reidy J. J., Beuerman R. W. Corneal nerve disruption reactivates virus in rabbits latently infected with HSV-1. Invest Ophthalmol Vis Sci. 1990 May;31(5):925–932. [PubMed] [Google Scholar]
- Bloom D. C., Devi-Rao G. B., Hill J. M., Stevens J. G., Wagner E. K. Molecular analysis of herpes simplex virus type 1 during epinephrine-induced reactivation of latently infected rabbits in vivo. J Virol. 1994 Mar;68(3):1283–1292. doi: 10.1128/jvi.68.3.1283-1292.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bourne N., Stanberry L. R., Connelly B. L., Kurawadwala J., Straus S. E., Krause P. R. Quantity of latency-associated transcript produced by herpes simplex virus is not predictive of the frequency of experimental recurrent genital herpes. J Infect Dis. 1994 May;169(5):1084–1087. doi: 10.1093/infdis/169.5.1084. [DOI] [PubMed] [Google Scholar]
- Cook S. D., Hill J. M., Lynas C., Maitland N. J. Latency-associated transcripts in corneas and ganglia of HSV-1 infected rabbits. Br J Ophthalmol. 1991 Nov;75(11):644–648. doi: 10.1136/bjo.75.11.644. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Demangone M., Hill J. M., Kwon B. S. Effects of acyclovir therapy during simultaneous reactivation of latent HSV-1 in rabbits. Antiviral Res. 1987 May;7(4):237–243. doi: 10.1016/0166-3542(87)90032-5. [DOI] [PubMed] [Google Scholar]
- Deshmane S. L., Fraser N. W. During latency, herpes simplex virus type 1 DNA is associated with nucleosomes in a chromatin structure. J Virol. 1989 Feb;63(2):943–947. doi: 10.1128/jvi.63.2.943-947.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Devi-Rao G. B., Bloom D. C., Stevens J. G., Wagner E. K. Herpes simplex virus type 1 DNA replication and gene expression during explant-induced reactivation of latently infected murine sensory ganglia. J Virol. 1994 Mar;68(3):1271–1282. doi: 10.1128/jvi.68.3.1271-1282.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ecob-Prince M. S., Hassan K., Denheen M. T., Preston C. M. Expression of beta-galactosidase in neurons of dorsal root ganglia which are latently infected with herpes simplex virus type 1. J Gen Virol. 1995 Jun;76(Pt 6):1527–1532. doi: 10.1099/0022-1317-76-6-1527. [DOI] [PubMed] [Google Scholar]
- Ecob-Prince M. S., Preston C. M., Rixon F. J., Hassan K., Kennedy P. G. 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 in1814. J Gen Virol. 1993 Jun;74(Pt 6):985–994. doi: 10.1099/0022-1317-74-6-985. [DOI] [PubMed] [Google Scholar]
- Ecob-Prince M. S., Rixon F. J., Preston C. M., Hassan K., Kennedy P. G. Reactivation in vivo and in vitro of herpes simplex virus from mouse dorsal root ganglia which contain different levels of latency-associated transcripts. J Gen Virol. 1993 Jun;74(Pt 6):995–1002. doi: 10.1099/0022-1317-74-6-995. [DOI] [PubMed] [Google Scholar]
- Farrell M. J., Hill J. M., Margolis T. P., Stevens J. G., Wagner E. K., Feldman L. T. The herpes simplex virus type 1 reactivation function lies outside the latency-associated transcript open reading frame ORF-2. J Virol. 1993 Jun;67(6):3653–3655. doi: 10.1128/jvi.67.6.3653-3655.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gressens P., Martin J. R. In situ polymerase chain reaction: localization of HSV-2 DNA sequences in infections of the nervous system. J Virol Methods. 1994 Jan;46(1):61–83. doi: 10.1016/0166-0934(94)90017-5. [DOI] [PubMed] [Google Scholar]
- Haruta Y., Maguire L. J., Rootman D. S., Hill J. M. Recurrent herpes simplex virus type 1 corneal epithelial lesions after radial keratotomy in the rabbit. Arch Ophthalmol. 1987 May;105(5):692–694. doi: 10.1001/archopht.1987.01060050110048. [DOI] [PubMed] [Google Scholar]
- Haruta Y., Rootman D. S., Hill J. M. Recurrent HSV-1 corneal epithelial lesions induced by timolol iontophoresis in latently infected rabbits. Invest Ophthalmol Vis Sci. 1988 Mar;29(3):387–392. [PubMed] [Google Scholar]
- Haruta Y., Rootman D. S., Xie L. X., Kiritoshi A., Hill J. M. Recurrent HSV-1 corneal lesions in rabbits induced by cyclophosphamide and dexamethasone. Invest Ophthalmol Vis Sci. 1989 Mar;30(3):371–376. [PubMed] [Google Scholar]
- Hill J. M., Haruta Y., Rootman D. S. Adrenergically induced recurrent HSV-1 corneal epithelial lesions. Curr Eye Res. 1987 Aug;6(8):1065–1071. doi: 10.3109/02713688709034878. [DOI] [PubMed] [Google Scholar]
- Hill J. M., Rayfield M. A., Haruta Y. Strain specificity of spontaneous and adrenergically induced HSV-1 ocular reactivation in latently infected rabbits. Curr Eye Res. 1987 Jan;6(1):91–97. doi: 10.3109/02713688709020074. [DOI] [PubMed] [Google Scholar]
- Hill J. M., Sedarati F., Javier R. T., Wagner E. K., Stevens J. G. Herpes simplex virus latent phase transcription facilitates in vivo reactivation. Virology. 1990 Jan;174(1):117–125. doi: 10.1016/0042-6822(90)90060-5. [DOI] [PubMed] [Google Scholar]
- Hill J. M., Shimomura Y., Kwon B. S., Gangarosa L. P., Sr Iontophoresis of epinephrine isomers to rabbit eyes induced HSV-1 ocular shedding. Invest Ophthalmol Vis Sci. 1985 Sep;26(9):1299–1303. [PubMed] [Google Scholar]
- Johnston R. F., Pickett S. C., Barker D. L. Autoradiography using storage phosphor technology. Electrophoresis. 1990 May;11(5):355–360. doi: 10.1002/elps.1150110503. [DOI] [PubMed] [Google Scholar]
- Katz J. P., Bodin E. T., Coen D. M. Quantitative polymerase chain reaction analysis of herpes simplex virus DNA in ganglia of mice infected with replication-incompetent mutants. J Virol. 1990 Sep;64(9):4288–4295. doi: 10.1128/jvi.64.9.4288-4295.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kramer M. F., Coen D. M. Quantification of transcripts from the ICP4 and thymidine kinase genes in mouse ganglia latently infected with herpes simplex virus. J Virol. 1995 Mar;69(3):1389–1399. doi: 10.1128/jvi.69.3.1389-1399.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kwon B. S., Gangarosa L. P., Burch K. D., deBack J., Hill J. M. Induction of ocular herpes simplex virus shedding by iontophoresis of epinephrine into rabbit cornea. Invest Ophthalmol Vis Sci. 1981 Sep;21(3):442–449. [PubMed] [Google Scholar]
- Lubin M. B., Elashoff J. D., Wang S. J., Rotter J. I., Toyoda H. Precise gene dosage determination by polymerase chain reaction: theory, methodology, and statistical approach. Mol Cell Probes. 1991 Aug;5(4):307–317. doi: 10.1016/0890-8508(91)90054-n. [DOI] [PubMed] [Google Scholar]
- Lynas C., Laycock K. A., Cook S. D., Hill T. J., Blyth W. A., Maitland N. J. Detection of herpes simplex virus type 1 gene expression in latently and productively infected mouse ganglia using the polymerase chain reaction. J Gen Virol. 1989 Sep;70(Pt 9):2345–2355. doi: 10.1099/0022-1317-70-9-2345. [DOI] [PubMed] [Google Scholar]
- Mehta A., Maggioncalda J., Bagasra O., Thikkavarapu S., Saikumari P., Valyi-Nagy T., Fraser N. W., Block T. M. In situ DNA PCR and RNA hybridization detection of herpes simplex virus sequences in trigeminal ganglia of latently infected mice. Virology. 1995 Jan 10;206(1):633–640. doi: 10.1016/s0042-6822(95)80080-8. [DOI] [PubMed] [Google Scholar]
- Perng G. C., Ghiasi H., Slanina S. M., Nesburn A. B., Wechsler S. L. The spontaneous reactivation function of the herpes simplex virus type 1 LAT gene resides completely within the first 1.5 kilobases of the 8.3-kilobase primary transcript. J Virol. 1996 Feb;70(2):976–984. doi: 10.1128/jvi.70.2.976-984.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ramakrishnan R., Levine M., Fink D. J. PCR-based analysis of herpes simplex virus type 1 latency in the rat trigeminal ganglion established with a ribonucleotide reductase-deficient mutant. J Virol. 1994 Nov;68(11):7083–7091. doi: 10.1128/jvi.68.11.7083-7091.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rock D. L., Fraser N. W. Detection of HSV-1 genome in central nervous system of latently infected mice. Nature. 1983 Apr 7;302(5908):523–525. doi: 10.1038/302523a0. [DOI] [PubMed] [Google Scholar]
- Rødahl E., Stevens J. G. Differential accumulation of herpes simplex virus type 1 latency-associated transcripts in sensory and autonomic ganglia. Virology. 1992 Jul;189(1):385–388. doi: 10.1016/0042-6822(92)90721-z. [DOI] [PubMed] [Google Scholar]
- Sandri-Goldin R. M., Levine M., Glorioso J. C. Method for induction of mutations in physically defined regions of the herpes simplex virus genome. J Virol. 1981 Apr;38(1):41–49. doi: 10.1128/jvi.38.1.41-49.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sawtell N. M., Thompson R. L. Herpes simplex virus type 1 latency-associated transcription unit promotes anatomical site-dependent establishment and reactivation from latency. J Virol. 1992 Apr;66(4):2157–2169. doi: 10.1128/jvi.66.4.2157-2169.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sedarati F., Izumi K. M., Wagner E. K., Stevens J. G. Herpes simplex virus type 1 latency-associated transcription plays no role in establishment or maintenance of a latent infection in murine sensory neurons. J Virol. 1989 Oct;63(10):4455–4458. doi: 10.1128/jvi.63.10.4455-4458.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stevens J. G., Wagner E. K., Devi-Rao G. B., Cook M. L., Feldman L. T. RNA complementary to a herpesvirus alpha gene mRNA is prominent in latently infected neurons. Science. 1987 Feb 27;235(4792):1056–1059. doi: 10.1126/science.2434993. [DOI] [PubMed] [Google Scholar]
- Stroop W. G., Rock D. L., Fraser N. W. Localization of herpes simplex virus in the trigeminal and olfactory systems of the mouse central nervous system during acute and latent infections by in situ hybridization. Lab Invest. 1984 Jul;51(1):27–38. [PubMed] [Google Scholar]
- Trousdale M. D., Steiner I., Spivack J. G., Deshmane S. L., Brown S. M., MacLean A. R., Subak-Sharpe J. H., Fraser N. W. In vivo and in vitro reactivation impairment of a herpes simplex virus type 1 latency-associated transcript variant in a rabbit eye model. J Virol. 1991 Dec;65(12):6989–6993. doi: 10.1128/jvi.65.12.6989-6993.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wagner E. K., Flanagan W. M., Devi-Rao G., Zhang Y. F., Hill J. M., Anderson K. P., Stevens J. G. The herpes simplex virus latency-associated transcript is spliced during the latent phase of infection. J Virol. 1988 Dec;62(12):4577–4585. doi: 10.1128/jvi.62.12.4577-4585.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]