1887

Abstract

Glycoprotein H (gH) of pseudorabies virus (PrV) is a structural component of the virion and forms a complex with another glycoprotein, gL. For a detailed analysis of the function of PrV gH, we isolated a gH-deficient mutant on trans-complementing gH-expressing cells after insertion of a β-galactosidase expression cassette into a partially deleted gH gene. The absence of gH did not affect primary or secondary attachment of PrV but the mutant was not infectious. The defect in infectivity could partially be overcome by experimentally induced membrane fusion using PEG, which suggests that gH was necessary for fusion between virion and cellular membranes. After intranasal inoculation into mice, the LD of complemented gH PrV was more than four orders of magnitude higher than that of wild-type PrV. Infection of the respiratory epithelium was much less efficient with complemented gH PrV as compared with rescued PrV, reflecting the lack of direct cell-to-cell spread. Complemented gH PrV was able to penetrate into a few trigeminal and sympathetic first order neurons accessible from the nasal cavity, whereas transneuronal transfer in the second order neurons was not observed. In summary, gH is essential for entry and cell-to-cell spread in cell culture, and for propagation in the nervous system of mice. This substantiates the hypothesis that transneuronal spread and direct cell-to-cell spread in cell culture are governed by similar mechanisms.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-77-9-2277
1996-09-01
2024-04-25
Loading full text...

Full text loading...

/deliver/fulltext/jgv/77/9/JV0770092277.html?itemId=/content/journal/jgv/10.1099/0022-1317-77-9-2277&mimeType=html&fmt=ahah

References

  1. Babic N., Mettenleiter T. C., Flamand A., Ugolini G. 1993; Role of essential glycoproteins gll and gp50 in transneuronal transfer of pseudorabies virus from the hypoglossal nerves of mice. Journal of Virology 67:4421–4426
    [Google Scholar]
  2. Babic N., Mettenleiter T. C., Ugolini G., Flamand A., Coulon P. 1994; Propagation of pseudorabies virus in the nervous system of the mouse after intranasal inoculation. Virology 204:616–625
    [Google Scholar]
  3. Babic N., Klupp B., Brack A., Mettenleiter T. C., Ugolini G., Flamand A. 1996; Deletion of glycoprotein gE reduces the propagation of pseudorabies virus in the nervous system of mice after intranasal inoculation. Virology 218: in press
    [Google Scholar]
  4. Balan P., Davis-Poynter N., Bell S., Atkinson S., Brown H., Minson T. 1994; An analysis of the in vitro and in vivo phenotypes of mutants of herpes simplex virus type 1 lacking glycoproteins gG, gE, gl or the putative gJ. Journal of General Virology 75:1245–1258
    [Google Scholar]
  5. Cai W., Gu B., Person S. 1988; Role of glycoprotein B of herpes simplex virus in viral entry and cell fusion. Journal of Virology 62:2596–2604
    [Google Scholar]
  6. Card J. P., Whealy M. E., Robbins A. K., Enquist L. W. 1992; Pseudorabies virus envelope glycoprotein gl influences both neurotropism and virulence during infection of the rat visual system. Journal of Neuroscience 66:3032–3041
    [Google Scholar]
  7. Coulon P., Derbin C., Kucera P., Lafay F., Prehaud C., Flamand A. 1989; Invasion of the peripheral nervous systems of adult mice by the CVS strain of rabies virus and its avirulent derivative AvO1. Journal of Virology 63:3550–3554
    [Google Scholar]
  8. Dingwell K. S., Doering L. C., Johnson D. C. 1995; Glycoproteins gE and gl facilitate neuron-to-neuron spread of herpes simplex virus. Journal of Virology 69:7087–7098
    [Google Scholar]
  9. Farrell H., McLean C., Harley C., Efstathiou S., Minson A. C. 1994; Vaccine potential of a herpes simplex virus type 1 mutant with an essential glycoprotein deleted. Journal of Virology 68:927–932
    [Google Scholar]
  10. Forrester A., Farrell H., Wilkinson G., Kaye J., Davis-Poynter N., Minson A. C. 1992; Construction and properties of a mutant of herpes simplex virus type 1 with glycoprotein H coding sequences deleted. Journal of Virology 66:341–348
    [Google Scholar]
  11. Graham F. L., van der Eb A. J. 1973; A new technique for the assay of infectivity of human adenovirus. Virology 52:456–467
    [Google Scholar]
  12. Heffner S., Kovacs F., Klupp B., Mettenleiter T. C. 1993; Glycoprotein gp50-negative pseudorabies virus: a novel approach toward a nonspreading live herpesvirus vaccine. Journal of Virology 67:1529–1537
    [Google Scholar]
  13. Kaplan A., Vatter A. 1959; A comparison of herpes simplex and pseudorabies viruses. Virology 7:394–407
    [Google Scholar]
  14. Karger A., Mettenleiter T. C. 1993; Glycoproteins gill and gp50 play dominant roles in the biphasic attachment of pseudorabies virus. Virology 194:654–664
    [Google Scholar]
  15. Klupp B., Visser N., Mettenleiter T. C. 1992; Identification and characterization of pseudorabies virus glycoprotein H. Journal of Virology 66:3048–3055
    [Google Scholar]
  16. Klupp B., Baumeister J., Karger A., Visser N., Mettenleiter T. C. 1994; Identification and characterization of a novel structural glycoprotein in pseudorabies virus, gL. Journal of Virology 68:3868–3878
    [Google Scholar]
  17. Kritas S. K., Pensaert M. B., Mettenleiter T. C. 1994; Role of envelope glycoproteins gl, gp63 and III in the invasion and spread of Aujeszky’s disease virus in the olfactory nervous system pathway of the pig. Journal of General Virology 75:2319–2327
    [Google Scholar]
  18. Lafay F., Coulon P., Astic L., Saucier D., Riche D., Holley A., Flamand A. 1991; Spread of the CVS strain of rabies virus and of the avirulent mutant AvO1 along the olfactory pathways of the mouse after intranasal inoculation. Virology 183:320–330
    [Google Scholar]
  19. Ligas M., Johnson D. 1988; A herpes simplex virus mutant in which glycoprotein D sequences are replaced by β-galactosidase sequences binds to but is unable to penetrate into cells. Journal of Virology 62:1486–1494
    [Google Scholar]
  20. Mettenleiter T. C. 1994; Pseudorabies (Aujeszky’s disease) virus: state of the art. Ada Veterinaria Hungarica 42:153–177
    [Google Scholar]
  21. Mettenleiter T. C., Rauh I. 1990; A glycoprotein gX-β-galactosidase fusion gene as insertional marker for rapid identification of pseudorabies mutants. Journal of Virological Methods 30:55–66
    [Google Scholar]
  22. Mettenleiter T. C., Zsak L., Zuckermann F., Sugg N., Kern H., Ben-Porat T. 1990; Interaction of glycoprotein gIII with a cellular heparinlike substance mediates adsorption of pseudorabies virus. Journal of Virology 64:278–286
    [Google Scholar]
  23. Mulder W. A. M., Jacobs L., Priem J., Kok G. L., Wagenaar F., Kimman T. G., Pol J. M. A. 1994; Glycoprotein E-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. Journal of General Virology 75:3095–3106
    [Google Scholar]
  24. Peeters B., Dewind N., Hooisma M., Wagenaar F., Gielkens A., Moormann R. 1992a; Pseudorabies virus envelope glycoproteins gp50 and g11 are essential for virus penetration but only g11 is involved in membrane fusion. Journal of Virology 66:894–905
    [Google Scholar]
  25. Peeters B., Dewind N., Broer R., Gielkens A., Moormann R. 1992b; Glycoprotein H of pseudorabies virus is essential for entry and cell-to-cell spread of the virus. Journal of Virology 66:3888–3892
    [Google Scholar]
  26. Peeters B., Pol J., Gielkens A., Moormann R. 1993; Envelope glycoprotein gp50 of pseudorabies virus is essential for virus entry but is not required for viral spread in mice. Journal of Virology 67:170–177
    [Google Scholar]
  27. Rauh I., Mettenleiter T. C. 1991; Pseudorabies virus glycoproteins gll and gp50 are essential for virus penetration. Journal of Virology 65:5348–5356
    [Google Scholar]
  28. Rauh I., Weiland F., Fehler F., Keil G., Mettenleiter T. C. 1991; Pseudorabies virus mutants lacking the essential glycoprotein g11 can be complemented by glycoprotein gl of bovine herpesvirus 1. Journal of Virology 65:621–631
    [Google Scholar]
  29. Sarmiento M., Haffey M., Spear P. G. 1979; Membrane proteins specified by herpes simplex viruses. III. Role of glycoprotein VP7(B2) in virion infectivity. Journal of Virology 29:1149–1158
    [Google Scholar]
  30. Sawitzky D., Hampl H., Habermehl K. O. 1990; Comparison of heparin-sensitive attachment of pseudorabies virus (PrV) and herpes simplex virus type 1 and identification of heparin-binding PrV glycoproteins. Journal of General Virology 71:1221–1225
    [Google Scholar]
  31. Southern P. J., Berg P. 1982; Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. Journal of Molecular and Applied Genetics 1:327–341
    [Google Scholar]
  32. Spear P. G. 1993; Entry of alphaherpesviruses into cells. Seminars in Virology 4:167–180
    [Google Scholar]
  33. Standish A., Enquist L. W., Schwaber J. S. 1994; Vagal cardiac ventricular innervation and its central medullary origin demonstrated by viral tracing. Science 263:232–234
    [Google Scholar]
  34. Strack A. M., Loewy A. D. 1990; Pseudorabies virus: a highly specific transneuronal cell body marker in the sympathetic nervous system. Journal of Neurosciences 10:2139–2147
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-77-9-2277
Loading
/content/journal/jgv/10.1099/0022-1317-77-9-2277
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error