Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1996 Apr;70(4):2191–2200. doi: 10.1128/jvi.70.4.2191-2200.1996

Glycoprotein D-negative pseudorabies virus can spread transneuronally via direct neuron-to-neuron transmission in its natural host, the pig, but not after additional inactivation of gE or gI.

W Mulder 1, J Pol 1, T Kimman 1, G Kok 1, J Priem 1, B Peeters 1
PMCID: PMC190058  PMID: 8642642

Abstract

Envelope glycoprotein D (gD) is essential for entry of pseudorabies virus (PRV) into cells but is not required for the subsequent steps in virus replication. Phenotypically complemented gD mutants can infect cells and can spread, both in vitro and in mice, by direct cell-to-cell transmission. Progeny virions released by infected cells are noninfectious because they lack gD. The aim of this study was to determine the role of gD in the neuropathogenicity of PRV in its natural host, the pig. We investigated whether gD-negative PRV can spread transneuronally via synaptically linked neurons of the olfactory and trigeminal routes. High doses of a phenotypically complemented gD mutant and gD mutants that are unable to express either gI or gI plus gE were inoculated intranasally in 3- to 5-week-old pigs. Compared with the wild-type virus, the virulence of the gD mutant was reduced. However, pigs inoculated with the gD mutant still developed fever and respiratory signs. Additional inactivation of either gI or gI plus gE further decreased virulence for pigs. Immunohistochemical examination of infected pigs showed that a PRV gD mutant could replicate and spread transneuronally into the central nervous system (CNS). Compared with the wild-type virus, the gD mutant had infected fewer neurons of the CNS on day 2. Nevertheless, on day 3, the gD-negative PRV had infected more neurons and viral antigens were present in second- and third-order neurons in the olfactory bulb, brain stem, and medulla oblongata. In contrast, gD mutants which are unable to express either gI or gI plus gE infected a limited number of first-order neurons in the olfactory epithelium and in the trigeminal ganglion and did not spread transneuronally or infect the CNS. Thus, transsynaptic spread of PRV in pigs can occur independently of gD. Possible mechanisms of transsynaptic transport of PRV are discussed.

Full Text

The Full Text of this article is available as a PDF (1.1 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Babic N., Mettenleiter T. C., Flamand A., Ugolini G. Role of essential glycoproteins gII and gp50 in transneuronal transfer of pseudorabies virus from the hypoglossal nerves of mice. J Virol. 1993 Jul;67(7):4421–4426. doi: 10.1128/jvi.67.7.4421-4426.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Card J. P., Rinaman L., Lynn R. B., Lee B. H., Meade R. P., Miselis R. R., Enquist L. W. Pseudorabies virus infection of the rat central nervous system: ultrastructural characterization of viral replication, transport, and pathogenesis. J Neurosci. 1993 Jun;13(6):2515–2539. doi: 10.1523/JNEUROSCI.13-06-02515.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. DeVries S. H., Baylor D. A. Synaptic circuitry of the retina and olfactory bulb. Cell. 1993 Jan;72 (Suppl):139–149. doi: 10.1016/s0092-8674(05)80033-9. [DOI] [PubMed] [Google Scholar]
  4. Eloit M., Fargeaud D., L'Haridon R., Toma B. Identification of the pseudorabies virus glycoprotein gp50 as a major target of neutralizing antibodies. Arch Virol. 1988;99(1-2):45–56. doi: 10.1007/BF01311022. [DOI] [PubMed] [Google Scholar]
  5. Enquist L. W., Dubin J., Whealy M. E., Card J. P. Complementation analysis of pseudorabies virus gE and gI mutants in retinal ganglion cell neurotropism. J Virol. 1994 Aug;68(8):5275–5279. doi: 10.1128/jvi.68.8.5275-5279.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Heffner S., Kovács F., Klupp B. G., Mettenleiter T. C. Glycoprotein gp50-negative pseudorabies virus: a novel approach toward a nonspreading live herpesvirus vaccine. J Virol. 1993 Mar;67(3):1529–1537. doi: 10.1128/jvi.67.3.1529-1537.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Jacobs L. Glycoprotein E of pseudorabies virus and homologous proteins in other alphaherpesvirinae. Arch Virol. 1994;137(3-4):209–228. doi: 10.1007/BF01309470. [DOI] [PubMed] [Google Scholar]
  8. Jacobs L., Mulder W. A., Van Oirschot J. T., Gielkens A. L., Kimman T. G. Deleting two amino acids in glycoprotein gI of pseudorabies virus decreases virulence and neurotropism for pigs, but does not affect immunogenicity. J Gen Virol. 1993 Oct;74(Pt 10):2201–2206. doi: 10.1099/0022-1317-74-10-2201. [DOI] [PubMed] [Google Scholar]
  9. Johnson D. C., Ligas M. W. Herpes simplex viruses lacking glycoprotein D are unable to inhibit virus penetration: quantitative evidence for virus-specific cell surface receptors. J Virol. 1988 Dec;62(12):4605–4612. doi: 10.1128/jvi.62.12.4605-4612.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Karger A., Mettenleiter T. C. Glycoproteins gIII and gp50 play dominant roles in the biphasic attachment of pseudorabies virus. Virology. 1993 Jun;194(2):654–664. doi: 10.1006/viro.1993.1305. [DOI] [PubMed] [Google Scholar]
  11. Kasza L., Shadduck J. A., Christofinis G. J. Establishment, viral susceptibility and biological characteristics of a swine kidney cell line SK-6. Res Vet Sci. 1972 Jan;13(1):46–51. [PubMed] [Google Scholar]
  12. Kimman T. G., De Wind N., De Bruin T., de Visser Y., Voermans J. Inactivation of glycoprotein gE and thymidine kinase or the US3-encoded protein kinase synergistically decreases in vivo replication of pseudorabies virus and the induction of protective immunity. Virology. 1994 Dec;205(2):511–518. doi: 10.1006/viro.1994.1672. [DOI] [PubMed] [Google Scholar]
  13. Kimman T. G., de Wind N., Oei-Lie N., Pol J. M., Berns A. J., Gielkens A. L. Contribution of single genes within the unique short region of Aujeszky's disease virus (suid herpesvirus type 1) to virulence, pathogenesis and immunogenicity. J Gen Virol. 1992 Feb;73(Pt 2):243–251. doi: 10.1099/0022-1317-73-2-243. [DOI] [PubMed] [Google Scholar]
  14. Kjellén L., Lindahl U. Proteoglycans: structures and interactions. Annu Rev Biochem. 1991;60:443–475. doi: 10.1146/annurev.bi.60.070191.002303. [DOI] [PubMed] [Google Scholar]
  15. Kost T. A., Jones E. V., Smith K. M., Reed A. P., Brown A. L., Miller T. J. Biological evaluation of glycoproteins mapping to two distinct mRNAs within the BamHI fragment 7 of pseudorabies virus: expression of the coding regions by vaccinia virus. Virology. 1989 Aug;171(2):365–376. doi: 10.1016/0042-6822(89)90604-1. [DOI] [PubMed] [Google Scholar]
  16. Kritas S. K., Pensaert M. B., Mettenleiter T. C. Invasion and spread of single glycoprotein deleted mutants of Aujeszky's disease virus (ADV) in the trigeminal nervous pathway of pigs after intranasal inoculation. Vet Microbiol. 1994 Jun;40(3-4):323–334. doi: 10.1016/0378-1135(94)90120-1. [DOI] [PubMed] [Google Scholar]
  17. Kritas S. K., Pensaert M. B., Mettenleiter T. C. Role of envelope glycoproteins gI, gp63 and gIII in the invasion and spread of Aujeszky's disease virus in the olfactory nervous pathway of the pig. J Gen Virol. 1994 Sep;75(Pt 9):2319–2327. doi: 10.1099/0022-1317-75-9-2319. [DOI] [PubMed] [Google Scholar]
  18. Kuypers H. G., Ugolini G. Viruses as transneuronal tracers. Trends Neurosci. 1990 Feb;13(2):71–75. doi: 10.1016/0166-2236(90)90071-h. [DOI] [PubMed] [Google Scholar]
  19. Martin X., Dolivo M. Neuronal and transneuronal tracing in the trigeminal system of the rat using the herpes virus suis. Brain Res. 1983 Aug 29;273(2):253–276. doi: 10.1016/0006-8993(83)90850-8. [DOI] [PubMed] [Google Scholar]
  20. McCracken R. M., McFerran J. B., Dow C. The neural spread of pseudorabies virus in calves. J Gen Virol. 1973 Jul;20(1):17–28. doi: 10.1099/0022-1317-20-1-17. [DOI] [PubMed] [Google Scholar]
  21. McFerran J. B., Dow C. Studies on immunisation of pigs with the Bartha strain of Aujeszky's disease virus. Res Vet Sci. 1975 Jul;19(1):17–22. [PubMed] [Google Scholar]
  22. Mettenleiter T. C. Initiation and spread of alpha-herpesvirus infections. Trends Microbiol. 1994 Jan;2(1):2–4. doi: 10.1016/0966-842x(94)90335-2. [DOI] [PubMed] [Google Scholar]
  23. Mettenleiter T. C., Klupp B. G., Weiland F., Visser N. Characterization of a quadruple glycoprotein-deleted pseudorabies virus mutant for use as a biologically safe live virus vaccine. J Gen Virol. 1994 Jul;75(Pt 7):1723–1733. doi: 10.1099/0022-1317-75-7-1723. [DOI] [PubMed] [Google Scholar]
  24. Mettenleiter T. C., Zsak L., Zuckermann F., Sugg N., Kern H., Ben-Porat T. Interaction of glycoprotein gIII with a cellular heparinlike substance mediates adsorption of pseudorabies virus. J Virol. 1990 Jan;64(1):278–286. doi: 10.1128/jvi.64.1.278-286.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Mulder W. A., Jacobs L., Priem J., Kok G. L., Wagenaar F., Kimman T. G., Pol J. M. Glycoprotein gE-negative pseudorabies virus has a reduced capability to infect second- and third-order neurons of the olfactory and trigeminal routes in the porcine central nervous system. J Gen Virol. 1994 Nov;75(Pt 11):3095–3106. doi: 10.1099/0022-1317-75-11-3095. [DOI] [PubMed] [Google Scholar]
  26. Mulder W. A., Priem J., Pol J. M., Kimman T. G. Role of viral proteins and concanavalin A in in vitro replication of pseudorabies virus in porcine peripheral blood mononuclear cells. J Gen Virol. 1995 Jun;76(Pt 6):1433–1442. doi: 10.1099/0022-1317-76-6-1433. [DOI] [PubMed] [Google Scholar]
  27. Peeters B., Bouma A., de Bruin T., Moormann R., Gielkens A., Kimman T. Non-transmissible pseudorabies virus gp50 mutants: a new generation of safe live vaccines. Vaccine. 1994 Mar;12(4):375–380. doi: 10.1016/0264-410x(94)90104-x. [DOI] [PubMed] [Google Scholar]
  28. Peeters B., Pol J., Gielkens A., Moormann R. Envelope glycoprotein gp50 of pseudorabies virus is essential for virus entry but is not required for viral spread in mice. J Virol. 1993 Jan;67(1):170–177. doi: 10.1128/jvi.67.1.170-177.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Peeters B., de Wind N., Broer R., Gielkens A., Moormann R. Glycoprotein H of pseudorabies virus is essential for entry and cell-to-cell spread of the virus. J Virol. 1992 Jun;66(6):3888–3892. doi: 10.1128/jvi.66.6.3888-3892.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Peeters B., de Wind N., Hooisma M., Wagenaar F., Gielkens A., Moormann R. Pseudorabies virus envelope glycoproteins gp50 and gII are essential for virus penetration, but only gII is involved in membrane fusion. J Virol. 1992 Feb;66(2):894–905. doi: 10.1128/jvi.66.2.894-905.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Petrovskis E. A., Timmins J. G., Armentrout M. A., Marchioli C. C., Yancey R. J., Jr, Post L. E. DNA sequence of the gene for pseudorabies virus gp50, a glycoprotein without N-linked glycosylation. J Virol. 1986 Aug;59(2):216–223. doi: 10.1128/jvi.59.2.216-223.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Pol J. M., Gielkens A. L., van Oirschot J. T. Comparative pathogenesis of three strains of pseudorabies virus in pigs. Microb Pathog. 1989 Nov;7(5):361–371. doi: 10.1016/0882-4010(89)90039-9. [DOI] [PubMed] [Google Scholar]
  33. Rauh I., Mettenleiter T. C. Pseudorabies virus glycoproteins gII and gp50 are essential for virus penetration. J Virol. 1991 Oct;65(10):5348–5356. doi: 10.1128/jvi.65.10.5348-5356.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Robbins A. K., Dorney D. J., Wathen M. W., Whealy M. E., Gold C., Watson R. J., Holland L. E., Weed S. D., Levine M., Glorioso J. C. The pseudorabies virus gII gene is closely related to the gB glycoprotein gene of herpes simplex virus. J Virol. 1987 Sep;61(9):2691–2701. doi: 10.1128/jvi.61.9.2691-2701.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Rziha H. J., Mettenleiter T. C., Ohlinger V., Wittmann G. Herpesvirus (pseudorabies virus) latency in swine: occurrence and physical state of viral DNA in neural tissues. Virology. 1986 Dec;155(2):600–613. doi: 10.1016/0042-6822(86)90220-5. [DOI] [PubMed] [Google Scholar]
  36. Sawitzky D., Hampl H., Habermehl K. O. Comparison of heparin-sensitive attachment of pseudorabies virus (PRV) and herpes simplex virus type 1 and identification of heparin-binding PRV glycoproteins. J Gen Virol. 1990 May;71(Pt 5):1221–1225. doi: 10.1099/0022-1317-71-5-1221. [DOI] [PubMed] [Google Scholar]
  37. Sears A. E., McGwire B. S., Roizman B. Infection of polarized MDCK cells with herpes simplex virus 1: two asymmetrically distributed cell receptors interact with different viral proteins. Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5087–5091. doi: 10.1073/pnas.88.12.5087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Strack A. M., Loewy A. D. Pseudorabies virus: a highly specific transneuronal cell body marker in the sympathetic nervous system. J Neurosci. 1990 Jul;10(7):2139–2147. doi: 10.1523/JNEUROSCI.10-07-02139.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wagenaar F., Pol J. M., Peeters B., Gielkens A. L., de Wind N., Kimman T. G. The US3-encoded protein kinase from pseudorabies virus affects egress of virions from the nucleus. J Gen Virol. 1995 Jul;76(Pt 7):1851–1859. doi: 10.1099/0022-1317-76-7-1851. [DOI] [PubMed] [Google Scholar]
  40. Zuckermann F. A., Mettenleiter T. C., Schreurs C., Sugg N., Ben-Porat T. Complex between glycoproteins gI and gp63 of pseudorabies virus: its effect on virus replication. J Virol. 1988 Dec;62(12):4622–4626. doi: 10.1128/jvi.62.12.4622-4626.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. de Wind N., Berns A., Gielkens A., Kimman T. Ribonucleotide reductase-deficient mutants of pseudorabies virus are avirulent for pigs and induce partial protective immunity. J Gen Virol. 1993 Mar;74(Pt 3):351–359. doi: 10.1099/0022-1317-74-3-351. [DOI] [PubMed] [Google Scholar]
  42. van Zijl M., Quint W., Briaire J., de Rover T., Gielkens A., Berns A. Regeneration of herpesviruses from molecularly cloned subgenomic fragments. J Virol. 1988 Jun;62(6):2191–2195. doi: 10.1128/jvi.62.6.2191-2195.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES