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. 1996 Sep;70(9):6519–6523. doi: 10.1128/jvi.70.9.6519-6523.1996

Detection of herpes simplex virus type 1 latency-associated transcript expression in trigeminal ganglia by in situ reverse transcriptase PCR.

R Ramakrishnan 1, P L Poliani 1, M Levine 1, J C Glorioso 1, D J Fink 1
PMCID: PMC190691  PMID: 8709293

Abstract

One of the defining characteristics of herpes simplex virus type 1 (HSV-1) infection is the ability of the virus to establish a lifelong latent state in neurons. We previously demonstrated (R. Ramakrishnan, A.J. Fink, G. Jiang, P. Desai, J. C. Glorioso, and M. Levine, J. Virol. 68:1864-1873, 1994) by in situ PCR that many more neurons contain viral genomes than are detected by in situ hybridization for HSV latency-associated transcripts (LATs). To determine whether all cells which contain genomes express LATs, we examined trigeminal ganglia for LATs 1 and 8 weeks after corneal scarification with ribonucleotide reductase-deficient HSV-1 by in situ reverse transcriptase PCR. The number of LAT-positive cells detected by in situ reverse transcriptase was substantially greater than the number of cells positive by in situ hybridization and appeared to be similar to the number of cells containing HSV genomes by in situ PCR and the number of ganglionic neurons that project to the cornea as detected by retrograde labeling with Fluorogold. These results demonstrate LAT expression in many neurons containing HSV-1 genomes.

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Selected References

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  1. Batchelor A. H., O'Hare P. Localization of cis-acting sequence requirements in the promoter of the latency-associated transcript of herpes simplex virus type 1 required for cell-type-specific activity. J Virol. 1992 Jun;66(6):3573–3582. doi: 10.1128/jvi.66.6.3573-3582.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Batchelor A. H., O'Hare P. Regulation and cell-type-specific activity of a promoter located upstream of the latency-associated transcript of herpes simplex virus type 1. J Virol. 1990 Jul;64(7):3269–3279. doi: 10.1128/jvi.64.7.3269-3279.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Batchelor A. H., Wilcox K. W., O'Hare P. Binding and repression of the latency-associated promoter of herpes simplex virus by the immediate early 175K protein. J Gen Virol. 1994 Apr;75(Pt 4):753–767. doi: 10.1099/0022-1317-75-4-753. [DOI] [PubMed] [Google Scholar]
  4. Chen X., Schmidt M. C., Goins W. F., Glorioso J. C. Two herpes simplex virus type 1 latency-active promoters differ in their contributions to latency-associated transcript expression during lytic and latent infections. J Virol. 1995 Dec;69(12):7899–7908. doi: 10.1128/jvi.69.12.7899-7908.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Coen D. M., Kosz-Vnenchak M., Jacobson J. G., Leib D. A., Bogard C. L., Schaffer P. A., Tyler K. L., Knipe D. M. Thymidine kinase-negative herpes simplex virus mutants establish latency in mouse trigeminal ganglia but do not reactivate. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4736–4740. doi: 10.1073/pnas.86.12.4736. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cook M. L., Bastone V. B., Stevens J. G. Evidence that neurons harbor latent herpes simplex virus. Infect Immun. 1974 May;9(5):946–951. doi: 10.1128/iai.9.5.946-951.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Deatly A. M., Spivack J. G., Lavi E., O'Boyle D. R., 2nd, Fraser N. W. Latent herpes simplex virus type 1 transcripts in peripheral and central nervous system tissues of mice map to similar regions of the viral genome. J Virol. 1988 Mar;62(3):749–756. doi: 10.1128/jvi.62.3.749-756.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Desai P., Ramakrishnan R., Lin Z. W., Osak B., Glorioso J. C., Levine M. The RR1 gene of herpes simplex virus type 1 is uniquely trans activated by ICP0 during infection. J Virol. 1993 Oct;67(10):6125–6135. doi: 10.1128/jvi.67.10.6125-6135.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Dobson A. T., Sederati F., Devi-Rao G., Flanagan W. M., Farrell M. J., Stevens J. G., Wagner E. K., Feldman L. T. Identification of the latency-associated transcript promoter by expression of rabbit beta-globin mRNA in mouse sensory nerve ganglia latently infected with a recombinant herpes simplex virus. J Virol. 1989 Sep;63(9):3844–3851. doi: 10.1128/jvi.63.9.3844-3851.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Efstathiou S., Minson A. C., Field H. J., Anderson J. R., Wildy P. Detection of herpes simplex virus-specific DNA sequences in latently infected mice and in humans. J Virol. 1986 Feb;57(2):446–455. doi: 10.1128/jvi.57.2.446-455.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fraser N. W., Block T. M., Spivack J. G. The latency-associated transcripts of herpes simplex virus: RNA in search of function. Virology. 1992 Nov;191(1):1–8. doi: 10.1016/0042-6822(92)90160-q. [DOI] [PubMed] [Google Scholar]
  13. Goins W. F., Sternberg L. R., Croen K. D., Krause P. R., Hendricks R. L., Fink D. J., Straus S. E., Levine M., Glorioso J. C. A novel latency-active promoter is contained within the herpes simplex virus type 1 UL flanking repeats. J Virol. 1994 Apr;68(4):2239–2252. doi: 10.1128/jvi.68.4.2239-2252.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jacobson J. G., Leib D. A., Goldstein D. J., Bogard C. L., Schaffer P. A., Weller S. K., Coen D. M. A herpes simplex virus ribonucleotide reductase deletion mutant is defective for productive acute and reactivatable latent infections of mice and for replication in mouse cells. Virology. 1989 Nov;173(1):276–283. doi: 10.1016/0042-6822(89)90244-4. [DOI] [PubMed] [Google Scholar]
  15. Kenny J. J., Krebs F. C., Hartle H. T., Gartner A. E., Chatton B., Leiden J. M., Hoeffler J. P., Weber P. C., Wigdahl B. Identification of a second ATF/CREB-like element in the herpes simplex virus type 1 (HSV-1) latency-associated transcript (LAT) promoter. Virology. 1994 Apr;200(1):220–235. doi: 10.1006/viro.1994.1180. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Mellerick D. M., Fraser N. W. Physical state of the latent herpes simplex virus genome in a mouse model system: evidence suggesting an episomal state. Virology. 1987 Jun;158(2):265–275. doi: 10.1016/0042-6822(87)90198-x. [DOI] [PubMed] [Google Scholar]
  18. Ramakrishnan R., Fink D. J., Jiang G., Desai P., Glorioso J. C., Levine M. Competitive quantitative PCR analysis of herpes simplex virus type 1 DNA and latency-associated transcript RNA in latently infected cells of the rat brain. J Virol. 1994 Mar;68(3):1864–1873. doi: 10.1128/jvi.68.3.1864-1873.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Rivera-Gonzalez R., Imbalzano A. N., Gu B., Deluca N. A. The role of ICP4 repressor activity in temporal expression of the IE-3 and latency-associated transcript promoters during HSV-1 infection. Virology. 1994 Aug 1;202(2):550–564. doi: 10.1006/viro.1994.1377. [DOI] [PubMed] [Google Scholar]
  21. Rock D. L., Nesburn A. B., Ghiasi H., Ong J., Lewis T. L., Lokensgard J. R., Wechsler S. L. Detection of latency-related viral RNAs in trigeminal ganglia of rabbits latently infected with herpes simplex virus type 1. J Virol. 1987 Dec;61(12):3820–3826. doi: 10.1128/jvi.61.12.3820-3826.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Stevens J. G. Human herpesviruses: a consideration of the latent state. Microbiol Rev. 1989 Sep;53(3):318–332. doi: 10.1128/mr.53.3.318-332.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. 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]
  25. Zwaagstra J. C., Ghiasi H., Nesburn A. B., Wechsler S. L. Identification of a major regulatory sequence in the latency associated transcript (LAT) promoter of herpes simplex virus type 1 (HSV-1). Virology. 1991 May;182(1):287–297. doi: 10.1016/0042-6822(91)90672-x. [DOI] [PubMed] [Google Scholar]

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