Abstract
PCR analysis of herpes simplex virus (HSV) genome replication and productive-cycle transcription was used to examine the role of the cornea in the latency-associated transcript (LAT)-mediated reactivation of HSV type 1 (HSV-1) in the rabbit eye model. The reduced relative reactivation frequency of 17 delta Pst (a LAT- virus) compared to those of wild-type and LAT+ rescuants correlated with reduced levels of viral DNA and transcription in the cornea following epinephrine induction. The timing of virus appearance in the cornea was most consistent with tissue peripheral to the cornea itself mediating a LAT-sensitive step in the reactivation process. Specific results include the following. (i) While viral DNA was found in the corneas of rabbits latently infected with either the LAT+ or LAT- virus prior to and during the first 16 to 24 h following induction, more was found in animals infected with the LAT+ virus. (ii) A significant increase in levels of viral DNA occurred 20 to 168 h following induction. (iii) The average relative amount of viral DNA was lower at all time points following reactivation of animals infected with the LAT- virus. (iv) Expression of productive-cycle transcripts could be detected in corneas of some rabbits latently infected with either the LAT+ or LAT- virus, and the amount recovered and the timing of appearance differed during the reactivation of rabbits latently infected with the LAT+ or LAT- virus. (v) Despite the reduced recoveries of LAT- virus DNA and productive-cycle transcripts in reactivating corneas in vivo compared to those of their LAT+ counterparts, such differences were not detected in cultured keratinocytes or in experiments in which relatively high titers of virus were superinfected into the eyes of latently infected rabbits. (vi) A number of LAT(+)-virus-infected rabbits expressed LAT in corneas isolated from uninduced rabbits. When seen, its amount was significantly higher than that of a productive-cycle (VP5) transcript.
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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]
- 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]
- Bloom D. C., Hill J. M., Devi-Rao G., Wagner E. K., Feldman L. T., Stevens J. G. A 348-base-pair region in the latency-associated transcript facilitates herpes simplex virus type 1 reactivation. J Virol. 1996 Apr;70(4):2449–2459. doi: 10.1128/jvi.70.4.2449-2459.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cook S. D., Batra S. K., Brown S. M. Recovery of herpes simplex virus from the corneas of experimentally infected rabbits. J Gen Virol. 1987 Jul;68(Pt 7):2013–2017. doi: 10.1099/0022-1317-68-7-2013. [DOI] [PubMed] [Google Scholar]
- Cook S. D., Hill J. H. Herpes simplex virus: molecular biology and the possibility of corneal latency. Surv Ophthalmol. 1991 Sep-Oct;36(2):140–148. doi: 10.1016/0039-6257(91)90127-2. [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]
- 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]
- Devi-Rao G. B., Goodart S. A., Hecht L. M., Rochford R., Rice M. K., Wagner E. K. Relationship between polyadenylated and nonpolyadenylated herpes simplex virus type 1 latency-associated transcripts. J Virol. 1991 May;65(5):2179–2190. doi: 10.1128/jvi.65.5.2179-2190.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gordon Y. J., Romanowski E., Araullo-Cruz T., McKnight J. L. HSV-1 corneal latency. Invest Ophthalmol Vis Sci. 1991 Mar;32(3):663–665. [PubMed] [Google Scholar]
- Hill J. M., Dudley J. B., Shimomura Y., Kaufman H. E. Quantitation and kinetics of induced HSV-1 ocular shedding. Curr Eye Res. 1986 Mar;5(3):241–246. doi: 10.3109/02713688609020049. [DOI] [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., Maggioncalda J. B., Garza H. H., Jr, Su Y. H., Fraser N. W., Block T. M. In vivo epinephrine reactivation of ocular herpes simplex virus type 1 in the rabbit is correlated to a 370-base-pair region located between the promoter and the 5' end of the 2.0 kilobase latency-associated transcript. J Virol. 1996 Oct;70(10):7270–7274. doi: 10.1128/jvi.70.10.7270-7274.1996. [DOI] [PMC free article] [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]
- Kaye S. B., Lynas C., Patterson A., Risk J. M., McCarthy K., Hart C. A. Evidence for herpes simplex viral latency in the human cornea. Br J Ophthalmol. 1991 Apr;75(4):195–200. doi: 10.1136/bjo.75.4.195. [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]
- Kwon B. S., Gangarosa L. P., Sr, Green K., Hill J. M. Kinetics of ocular herpes simplex virus shedding induced by epinephrine iontophoresis. Invest Ophthalmol Vis Sci. 1982 Jun;22(6):818–821. [PubMed] [Google Scholar]
- Maggioncalda J., Mehta A., Fraser N. W., Block T. M. Analysis of a herpes simplex virus type 1 LAT mutant with a deletion between the putative promoter and the 5' end of the 2.0-kilobase transcript. J Virol. 1994 Dec;68(12):7816–7824. doi: 10.1128/jvi.68.12.7816-7824.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maggioncalda J., Mehta A., Su Y. H., Fraser N. W., Block T. M. Correlation between herpes simplex virus type 1 rate of reactivation from latent infection and the number of infected neurons in trigeminal ganglia. Virology. 1996 Nov 1;225(1):72–81. doi: 10.1006/viro.1996.0576. [DOI] [PubMed] [Google Scholar]
- Morris D. J., Cleator G. M., Klapper P. E., Cooper R. J., Biney E. O., Dennett C., Marcyniuk B., Tullo A. B. Detection of herpes simplex virus DNA in donor cornea culture medium by polymerase chain reaction. Br J Ophthalmol. 1996 Jul;80(7):654–657. doi: 10.1136/bjo.80.7.654. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Openshaw H., McNeill J. I., Lin X. H., Niland J., Cantin E. M. Herpes simplex virus DNA in normal corneas: persistence without viral shedding from ganglia. J Med Virol. 1995 May;46(1):75–80. doi: 10.1002/jmv.1890460116. [DOI] [PubMed] [Google Scholar]
- Perng G. C., Dunkel E. C., Geary P. A., Slanina S. M., Ghiasi H., Kaiwar R., Nesburn A. B., Wechsler S. L. The latency-associated transcript gene of herpes simplex virus type 1 (HSV-1) is required for efficient in vivo spontaneous reactivation of HSV-1 from latency. J Virol. 1994 Dec;68(12):8045–8055. doi: 10.1128/jvi.68.12.8045-8055.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rock D. L., Hagemoser W. A., Osorio F. A., Reed D. E. Detection of bovine herpesvirus type 1 RNA in trigeminal ganglia of latently infected rabbits by in situ hybridization. J Gen Virol. 1986 Nov;67(Pt 11):2515–2520. doi: 10.1099/0022-1317-67-11-2515. [DOI] [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]
- Sawtell N. M., Thompson R. L. Rapid in vivo reactivation of herpes simplex virus in latently infected murine ganglionic neurons after transient hyperthermia. J Virol. 1992 Apr;66(4):2150–2156. doi: 10.1128/jvi.66.4.2150-2156.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sedarati F., Margolis T. P., Stevens J. G. Latent infection can be established with drastically restricted transcription and replication of the HSV-1 genome. Virology. 1993 Feb;192(2):687–691. doi: 10.1006/viro.1993.1089. [DOI] [PubMed] [Google Scholar]
- Singer-Sam J., Robinson M. O., Bellvé A. R., Simon M. I., Riggs A. D. Measurement by quantitative PCR of changes in HPRT, PGK-1, PGK-2, APRT, MTase, and Zfy gene transcripts during mouse spermatogenesis. Nucleic Acids Res. 1990 Mar 11;18(5):1255–1259. doi: 10.1093/nar/18.5.1255. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thompson R. L., Wagner E. K. Partial rescue of herpes simplex virus neurovirulence with a 3.2 kb cloned DNA fragment. Virus Genes. 1988 Jun;1(3):261–273. doi: 10.1007/BF00572705. [DOI] [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., Bloom D. C. Experimental investigation of herpes simplex virus latency. Clin Microbiol Rev. 1997 Jul;10(3):419–443. doi: 10.1128/cmr.10.3.419. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wagner E. K., Devi-Rao G., Feldman L. T., Dobson A. T., Zhang Y. F., Flanagan W. M., Stevens J. G. Physical characterization of the herpes simplex virus latency-associated transcript in neurons. J Virol. 1988 Apr;62(4):1194–1202. doi: 10.1128/jvi.62.4.1194-1202.1988. [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]
- Xie L., Gebhardt B. M. A simplified technique for the short-term tissue culture of rabbit corneal cells. In Vitro Cell Dev Biol. 1989 Jan;25(1):20–22. doi: 10.1007/BF02624406. [DOI] [PubMed] [Google Scholar]