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. 1995 Aug;69(8):4593–4599. doi: 10.1128/jvi.69.8.4593-4599.1995

Long-term expression in sensory neurons in tissue culture from herpes simplex virus type 1 (HSV-1) promoters in an HSV-1-derived vector.

R L Smith 1, A I Geller 1, K W Escudero 1, C L Wilcox 1
PMCID: PMC189257  PMID: 7609023

Abstract

Amplicons, defective herpes simplex virus type 1 (HSV-1) vectors, were constructed to use four HSV-1 promoters, from the immediate-early (IE) 1 IE 3, IE 4/5, and late glycoprotein C (gC) genes, to regulate expression of the Escherichia coli lacZ gene, encoding beta-galactosidase, and packaged into infectious particles. Infection of sensory neurons in vitro with amplicons containing the IE 1, IE 3, or IE 4/5 promoter resulted in stable long-term expression of beta-galactosidase from 2 to 10 weeks after gene transfer. The number of neurons expressing beta-galactosidase was not changed by treatments previously shown to produce reactivation of latent HSV-1. In addition, the latency-associated transcript was detected in many of the same neurons that expressed beta-galactosidase, indicating that the viral IE promoters in the amplicons can function in the same neurons that harbor latent virus. Delivery of beta-galactosidase protein directly into neurons by microinjection indicated that the half-life for histochemical detection of beta-galactosidase was between 24 and 48 h, suggesting that the persistence of beta-galactosidase histochemical staining cannot be explained by the stability of the reporter protein alone. In contrast to the IE promoters, the gC promoter of the late gene class did not support long-term expression of beta-galactosidase; instead, beta-galactosidase was detected in only a few neurons per culture at 2 weeks after infection, and superinfection with wild-type HSV-1 did not increase the level of expression from the gC promoter. These results suggest that the HSV-1 IE promoters in the amplicons are not subject to the promoter inactivation that occurs with many types of virus vectors and that the IE promoters in the context of the amplicon avoid the promoter inactivation observed from the same promoters in the HSV-1 genome during latency.

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Selected References

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  1. Arthur J., Efstathiou S., Simmons A. Intranuclear foci containing low abundance herpes simplex virus latency-associated transcripts visualized by non-isotopic in situ hybridization. J Gen Virol. 1993 Jul;74(Pt 7):1363–1370. doi: 10.1099/0022-1317-74-7-1363. [DOI] [PubMed] [Google Scholar]
  2. Battleman D. S., Geller A. I., Chao M. V. HSV-1 vector-mediated gene transfer of the human nerve growth factor receptor p75hNGFR defines high-affinity NGF binding. J Neurosci. 1993 Mar;13(3):941–951. doi: 10.1523/JNEUROSCI.13-03-00941.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bergold P. J., Casaccia-Bonnefil P., Zeng X. L., Federoff H. J. Transsynaptic neuronal loss induced in hippocampal slice cultures by a herpes simplex virus vector expressing the GluR6 subunit of the kainate receptor. Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):6165–6169. doi: 10.1073/pnas.90.13.6165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chiocca E. A., Choi B. B., Cai W. Z., DeLuca N. A., Schaffer P. A., DiFiglia M., Breakefield X. O., Martuza R. L. Transfer and expression of the lacZ gene in rat brain neurons mediated by herpes simplex virus mutants. New Biol. 1990 Aug;2(8):739–746. [PubMed] [Google Scholar]
  5. Deatly A. M., Spivack J. G., Lavi E., Fraser N. W. 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 U S A. 1987 May;84(10):3204–3208. doi: 10.1073/pnas.84.10.3204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dobson A. T., Margolis T. P., Sedarati F., Stevens J. G., Feldman L. T. A latent, nonpathogenic HSV-1-derived vector stably expresses beta-galactosidase in mouse neurons. Neuron. 1990 Sep;5(3):353–360. doi: 10.1016/0896-6273(90)90171-b. [DOI] [PubMed] [Google Scholar]
  7. During M. J., Naegele J. R., O'Malley K. L., Geller A. I. Long-term behavioral recovery in parkinsonian rats by an HSV vector expressing tyrosine hydroxylase. Science. 1994 Nov 25;266(5189):1399–1403. doi: 10.1126/science.266.5189.1399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Federoff H. J., Geschwind M. D., Geller A. I., Kessler J. A. Expression of nerve growth factor in vivo from a defective herpes simplex virus 1 vector prevents effects of axotomy on sympathetic ganglia. Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1636–1640. doi: 10.1073/pnas.89.5.1636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Geller A. I. A new method to propagate defective HSV-1 vectors. Nucleic Acids Res. 1988 Jun 24;16(12):5690–5690. doi: 10.1093/nar/16.12.5690. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Geller A. I., Breakefield X. O. A defective HSV-1 vector expresses Escherichia coli beta-galactosidase in cultured peripheral neurons. Science. 1988 Sep 23;241(4873):1667–1669. doi: 10.1126/science.241.4873.1667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Geller A. I., During M. J., Haycock J. W., Freese A., Neve R. Long-term increases in neurotransmitter release from neuronal cells expressing a constitutively active adenylate cyclase from a herpes simplex virus type 1 vector. Proc Natl Acad Sci U S A. 1993 Aug 15;90(16):7603–7607. doi: 10.1073/pnas.90.16.7603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Geller A. I., During M. J., Oh Y. J., Freese A., O'Malley K. An HSV-1 vector expressing tyrosine hydroxylase causes production and release of L-dopa from cultured rat striatal cells. J Neurochem. 1995 Feb;64(2):487–496. doi: 10.1046/j.1471-4159.1995.64020487.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Geller A. I., Freese A. Infection of cultured central nervous system neurons with a defective herpes simplex virus 1 vector results in stable expression of Escherichia coli beta-galactosidase. Proc Natl Acad Sci U S A. 1990 Feb;87(3):1149–1153. doi: 10.1073/pnas.87.3.1149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Geschwind M. D., Kessler J. A., Geller A. I., Federoff H. J. Transfer of the nerve growth factor gene into cell lines and cultured neurons using a defective herpes simplex virus vector. Transfer of the NGF gene into cells by a HSV-1 vector. Brain Res Mol Brain Res. 1994 Jul;24(1-4):327–335. doi: 10.1016/0169-328x(94)90146-5. [DOI] [PubMed] [Google Scholar]
  16. Ho D. Y., Mocarski E. S., Sapolsky R. M. Altering central nervous system physiology with a defective herpes simplex virus vector expressing the glucose transporter gene. Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3655–3659. doi: 10.1073/pnas.90.8.3655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kaplitt M. G., Kwong A. D., Kleopoulos S. P., Mobbs C. V., Rabkin S. D., Pfaff D. W. Preproenkephalin promoter yields region-specific and long-term expression in adult brain after direct in vivo gene transfer via a defective herpes simplex viral vector. Proc Natl Acad Sci U S A. 1994 Sep 13;91(19):8979–8983. doi: 10.1073/pnas.91.19.8979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Kwong A. D., Frenkel N. Herpes simplex virus amplicon: effect of size on replication of constructed defective genomes containing eucaryotic DNA sequences. J Virol. 1984 Sep;51(3):595–603. doi: 10.1128/jvi.51.3.595-603.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lillycrop K. A., Dawson S. J., Estridge J. K., Gerster T., Matthias P., Latchman D. S. Repression of a herpes simplex virus immediate-early promoter by the Oct-2 transcription factor is dependent on an inhibitory region at the N terminus of the protein. Mol Cell Biol. 1994 Nov;14(11):7633–7642. doi: 10.1128/mcb.14.11.7633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lokensgard J. R., Bloom D. C., Dobson A. T., Feldman L. T. Long-term promoter activity during herpes simplex virus latency. J Virol. 1994 Nov;68(11):7148–7158. doi: 10.1128/jvi.68.11.7148-7158.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. McGeoch D. J., Dalrymple M. A., Davison A. J., Dolan A., Frame M. C., McNab D., Perry L. J., Scott J. E., Taylor P. The complete DNA sequence of the long unique region in the genome of herpes simplex virus type 1. J Gen Virol. 1988 Jul;69(Pt 7):1531–1574. doi: 10.1099/0022-1317-69-7-1531. [DOI] [PubMed] [Google Scholar]
  23. McGeoch D. J. The genomes of the human herpesviruses: contents, relationships, and evolution. Annu Rev Microbiol. 1989;43:235–265. doi: 10.1146/annurev.mi.43.100189.001315. [DOI] [PubMed] [Google Scholar]
  24. Paterson T., Everett R. D. A prominent serine-rich region in Vmw175, the major transcriptional regulator protein of herpes simplex virus type 1, is not essential for virus growth in tissue culture. J Gen Virol. 1990 Aug;71(Pt 8):1775–1783. doi: 10.1099/0022-1317-71-8-1775. [DOI] [PubMed] [Google Scholar]
  25. Sanes J. R., Rubenstein J. L., Nicolas J. F. Use of a recombinant retrovirus to study post-implantation cell lineage in mouse embryos. EMBO J. 1986 Dec 1;5(12):3133–3142. doi: 10.1002/j.1460-2075.1986.tb04620.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schmued L., Kyriakidis K., Heimer L. In vivo anterograde and retrograde axonal transport of the fluorescent rhodamine-dextran-amine, Fluoro-Ruby, within the CNS. Brain Res. 1990 Aug 27;526(1):127–134. doi: 10.1016/0006-8993(90)90258-d. [DOI] [PubMed] [Google Scholar]
  27. Smith R. L., Escudero J. M., Wilcox C. L. Regulation of the herpes simplex virus latency-associated transcripts during establishment of latency in sensory neurons in vitro. Virology. 1994 Jul;202(1):49–60. doi: 10.1006/viro.1994.1321. [DOI] [PubMed] [Google Scholar]
  28. Smith R. L., Pizer L. I., Johnson E. M., Jr, Wilcox C. L. Activation of second-messenger pathways reactivates latent herpes simplex virus in neuronal cultures. Virology. 1992 May;188(1):311–318. doi: 10.1016/0042-6822(92)90760-m. [DOI] [PubMed] [Google Scholar]
  29. Spaete R. R., Frenkel N. The herpes simplex virus amplicon: a new eucaryotic defective-virus cloning-amplifying vector. Cell. 1982 Aug;30(1):295–304. doi: 10.1016/0092-8674(82)90035-6. [DOI] [PubMed] [Google Scholar]
  30. Spaete R. R., Mocarski E. S. Insertion and deletion mutagenesis of the human cytomegalovirus genome. Proc Natl Acad Sci U S A. 1987 Oct;84(20):7213–7217. doi: 10.1073/pnas.84.20.7213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Stevens J. G. Latent herpes simplex virus and the nervous system,. Curr Top Microbiol Immunol. 1975;70:31–50. doi: 10.1007/978-3-642-66101-3_2. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Wilcox C. L., Johnson E. M., Jr Nerve growth factor deprivation results in the reactivation of latent herpes simplex virus in vitro. J Virol. 1987 Jul;61(7):2311–2315. doi: 10.1128/jvi.61.7.2311-2315.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wilcox C. L., Smith R. L., Freed C. R., Johnson E. M., Jr Nerve growth factor-dependence of herpes simplex virus latency in peripheral sympathetic and sensory neurons in vitro. J Neurosci. 1990 Apr;10(4):1268–1275. doi: 10.1523/JNEUROSCI.10-04-01268.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]

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