Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1997 Aug;71(8):5805–5813. doi: 10.1128/jvi.71.8.5805-5813.1997

Characterization of a BHK(TK-) cell clone resistant to postattachment entry by herpes simplex virus types 1 and 2.

R J Roller 1, B C Herold 1
PMCID: PMC191835  PMID: 9223469

Abstract

BHK(TK-) cells selected for resistance to polyethylene glycol-mediated fusion give rise to clones that are resistant to herpes simplex virus (HSV) infection. We have characterized one such clone, designated 95-19, and found that it is resistant to entry of HSV type 1 (HSV-1), HSV-2, and the related alphaherpesvirus pseudorabies virus (PRV). Single-step growth experiments show no detectable replication of multiple strains of HSV-1 and HSV-2 on 95-19 cells. Three lines of evidence suggest that these cells are resistant to postattachment entry. (i) Measurements of binding of radiolabeled virus show that heparin-sensitive binding of HSV-1 and HSV-2 to 95-19 cells is identical to binding to BHK(TK-) cells, suggesting that the block to replication occurs after attachment to heparan sulfate proteoglycan. (ii) 95-19 cells exposed to HSV-1 or HSV-2 at high multiplicity show no detectable immediate-early (IE) mRNA expression. (iii) Exposure of attached virus and cells to polyethylene glycol results in partial recovery of both IE gene expression and virus yield in single-step growth. The degrees of recovery of single-step yield and IE gene expression are similar, suggesting that the only block to single-step replication is at the point of virus entry and that these cells are deficient in some cellular factor required for efficient postattachment entry of free virus. 95-19 cells are also highly resistant to entry by cell-to-cell spread, suggesting that the same cellular factor participates in both types of entry.

Full Text

The Full Text of this article is available as a PDF (349.0 KB).

Selected References

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

  1. Balan P., Davis-Poynter N., Bell S., Atkinson H., Browne H., Minson T. An analysis of the in vitro and in vivo phenotypes of mutants of herpes simplex virus type 1 lacking glycoproteins gG, gE, gI or the putative gJ. J Gen Virol. 1994 Jun;75(Pt 6):1245–1258. doi: 10.1099/0022-1317-75-6-1245. [DOI] [PubMed] [Google Scholar]
  2. Brandimarti R., Huang T., Roizman B., Campadelli-Fiume G. Mapping of herpes simplex virus 1 genes with mutations which overcome host restrictions to infection. Proc Natl Acad Sci U S A. 1994 Jun 7;91(12):5406–5410. doi: 10.1073/pnas.91.12.5406. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brunetti C. R., Burke R. L., Hoflack B., Ludwig T., Dingwell K. S., Johnson D. C. Role of mannose-6-phosphate receptors in herpes simplex virus entry into cells and cell-to-cell transmission. J Virol. 1995 Jun;69(6):3517–3528. doi: 10.1128/jvi.69.6.3517-3528.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brunetti C. R., Burke R. L., Kornfeld S., Gregory W., Masiarz F. R., Dingwell K. S., Johnson D. C. Herpes simplex virus glycoprotein D acquires mannose 6-phosphate residues and binds to mannose 6-phosphate receptors. J Biol Chem. 1994 Jun 24;269(25):17067–17074. [PubMed] [Google Scholar]
  5. Cai W. H., Gu B., Person S. Role of glycoprotein B of herpes simplex virus type 1 in viral entry and cell fusion. J Virol. 1988 Aug;62(8):2596–2604. doi: 10.1128/jvi.62.8.2596-2604.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Campadelli-Fiume G., Arsenakis M., Farabegoli F., Roizman B. Entry of herpes simplex virus 1 in BJ cells that constitutively express viral glycoprotein D is by endocytosis and results in degradation of the virus. J Virol. 1988 Jan;62(1):159–167. doi: 10.1128/jvi.62.1.159-167.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Campadelli-Fiume G., Qi S., Avitabile E., Foà-Tomasi L., Brandimarti R., Roizman B. Glycoprotein D of herpes simplex virus encodes a domain which precludes penetration of cells expressing the glycoprotein by superinfecting herpes simplex virus. J Virol. 1990 Dec;64(12):6070–6079. doi: 10.1128/jvi.64.12.6070-6079.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dean H. J., Terhune S. S., Shieh M. T., Susmarski N., Spear P. G. Single amino acid substitutions in gD of herpes simplex virus 1 confer resistance to gD-mediated interference and cause cell-type-dependent alterations in infectivity. Virology. 1994 Feb 15;199(1):67–80. doi: 10.1006/viro.1994.1098. [DOI] [PubMed] [Google Scholar]
  9. Dingwell K. S., Brunetti C. R., Hendricks R. L., Tang Q., Tang M., Rainbow A. J., Johnson D. C. Herpes simplex virus glycoproteins E and I facilitate cell-to-cell spread in vivo and across junctions of cultured cells. J Virol. 1994 Feb;68(2):834–845. doi: 10.1128/jvi.68.2.834-845.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dingwell K. S., Doering L. C., Johnson D. C. Glycoproteins E and I facilitate neuron-to-neuron spread of herpes simplex virus. J Virol. 1995 Nov;69(11):7087–7098. doi: 10.1128/jvi.69.11.7087-7098.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Forrester A., Farrell H., Wilkinson G., Kaye J., Davis-Poynter N., Minson T. Construction and properties of a mutant of herpes simplex virus type 1 with glycoprotein H coding sequences deleted. J Virol. 1992 Jan;66(1):341–348. doi: 10.1128/jvi.66.1.341-348.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fuller A. O., Lee W. C. Herpes simplex virus type 1 entry through a cascade of virus-cell interactions requires different roles of gD and gH in penetration. J Virol. 1992 Aug;66(8):5002–5012. doi: 10.1128/jvi.66.8.5002-5012.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fuller A. O., Spear P. G. Anti-glycoprotein D antibodies that permit adsorption but block infection by herpes simplex virus 1 prevent virion-cell fusion at the cell surface. Proc Natl Acad Sci U S A. 1987 Aug;84(15):5454–5458. doi: 10.1073/pnas.84.15.5454. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gruenheid S., Gatzke L., Meadows H., Tufaro F. Herpes simplex virus infection and propagation in a mouse L cell mutant lacking heparan sulfate proteoglycans. J Virol. 1993 Jan;67(1):93–100. doi: 10.1128/jvi.67.1.93-100.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Herold B. C., Gerber S. I., Belval B. J., Siston A. M., Shulman N. Differences in the susceptibility of herpes simplex virus types 1 and 2 to modified heparin compounds suggest serotype differences in viral entry. J Virol. 1996 Jun;70(6):3461–3469. doi: 10.1128/jvi.70.6.3461-3469.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Herold B. C., Visalli R. J., Susmarski N., Brandt C. R., Spear P. G. Glycoprotein C-independent binding of herpes simplex virus to cells requires cell surface heparan sulphate and glycoprotein B. J Gen Virol. 1994 Jun;75(Pt 6):1211–1222. doi: 10.1099/0022-1317-75-6-1211. [DOI] [PubMed] [Google Scholar]
  17. Herold B. C., WuDunn D., Soltys N., Spear P. G. Glycoprotein C of herpes simplex virus type 1 plays a principal role in the adsorption of virus to cells and in infectivity. J Virol. 1991 Mar;65(3):1090–1098. doi: 10.1128/jvi.65.3.1090-1098.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Highlander S. L., Sutherland S. L., Gage P. J., Johnson D. C., Levine M., Glorioso J. C. Neutralizing monoclonal antibodies specific for herpes simplex virus glycoprotein D inhibit virus penetration. J Virol. 1987 Nov;61(11):3356–3364. doi: 10.1128/jvi.61.11.3356-3364.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hutchinson L., Graham F. L., Cai W., Debroy C., Person S., Johnson D. C. Herpes simplex virus (HSV) glycoproteins B and K inhibit cell fusion induced by HSV syncytial mutants. Virology. 1993 Oct;196(2):514–531. doi: 10.1006/viro.1993.1507. [DOI] [PubMed] [Google Scholar]
  20. Johnson D. C., Burke R. L., Gregory T. Soluble forms of herpes simplex virus glycoprotein D bind to a limited number of cell surface receptors and inhibit virus entry into cells. J Virol. 1990 Jun;64(6):2569–2576. doi: 10.1128/jvi.64.6.2569-2576.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Johnson R. M., Spear P. G. Herpes simplex virus glycoprotein D mediates interference with herpes simplex virus infection. J Virol. 1989 Feb;63(2):819–827. doi: 10.1128/jvi.63.2.819-827.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Jüttermann R., Weyer U., Doerfler W. Defect of adenovirus type 12 replication in hamster cells: absence of transcription of viral virus-associated and L1 RNAs. J Virol. 1989 Aug;63(8):3535–3540. doi: 10.1128/jvi.63.8.3535-3540.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Lee W. C., Fuller A. O. Herpes simplex virus type 1 and pseudorabies virus bind to a common saturable receptor on Vero cells that is not heparan sulfate. J Virol. 1993 Sep;67(9):5088–5097. doi: 10.1128/jvi.67.9.5088-5097.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Ligas M. W., Johnson D. C. A herpes simplex virus mutant in which glycoprotein D sequences are replaced by beta-galactosidase sequences binds to but is unable to penetrate into cells. J Virol. 1988 May;62(5):1486–1494. doi: 10.1128/jvi.62.5.1486-1494.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. MacLean C. A., Efstathiou S., Elliott M. L., Jamieson F. E., McGeoch D. J. Investigation of herpes simplex virus type 1 genes encoding multiply inserted membrane proteins. J Gen Virol. 1991 Apr;72(Pt 4):897–906. doi: 10.1099/0022-1317-72-4-897. [DOI] [PubMed] [Google Scholar]
  28. Matlin K. S., Reggio H., Helenius A., Simons K. Pathway of vesicular stomatitis virus entry leading to infection. J Mol Biol. 1982 Apr 15;156(3):609–631. doi: 10.1016/0022-2836(82)90269-8. [DOI] [PubMed] [Google Scholar]
  29. Montgomery R. I., Warner M. S., Lum B. J., Spear P. G. Herpes simplex virus-1 entry into cells mediated by a novel member of the TNF/NGF receptor family. Cell. 1996 Nov 1;87(3):427–436. doi: 10.1016/s0092-8674(00)81363-x. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. 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]
  32. 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]
  33. Roller R. J., Roizman B. A herpes simplex virus 1 US11-expressing cell line is resistant to herpes simplex virus infection at a step in viral entry mediated by glycoprotein D. J Virol. 1994 May;68(5):2830–2839. doi: 10.1128/jvi.68.5.2830-2839.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Roop C., Hutchinson L., Johnson D. C. A mutant herpes simplex virus type 1 unable to express glycoprotein L cannot enter cells, and its particles lack glycoprotein H. J Virol. 1993 Apr;67(4):2285–2297. doi: 10.1128/jvi.67.4.2285-2297.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sarmiento M., Haffey M., Spear P. G. Membrane proteins specified by herpes simplex viruses. III. Role of glycoprotein VP7(B2) in virion infectivity. J Virol. 1979 Mar;29(3):1149–1158. doi: 10.1128/jvi.29.3.1149-1158.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. 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]
  37. Shieh M. T., Spear P. G. Herpesvirus-induced cell fusion that is dependent on cell surface heparan sulfate or soluble heparin. J Virol. 1994 Feb;68(2):1224–1228. doi: 10.1128/jvi.68.2.1224-1228.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Shieh M. T., WuDunn D., Montgomery R. I., Esko J. D., Spear P. G. Cell surface receptors for herpes simplex virus are heparan sulfate proteoglycans. J Cell Biol. 1992 Mar;116(5):1273–1281. doi: 10.1083/jcb.116.5.1273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Singh J., Wagner E. K. Herpes simplex virus recombination vectors designed to allow insertion of modified promoters into transcriptionally "neutral" segments of the viral genome. Virus Genes. 1995;10(2):127–136. doi: 10.1007/BF01702593. [DOI] [PubMed] [Google Scholar]
  40. Subramanian G., LeBlanc R. A., Wardley R. C., Fuller A. O. Defective entry of herpes simplex virus types 1 and 2 into porcine cells and lack of infection in infant pigs indicate species tropism. J Gen Virol. 1995 Sep;76(Pt 9):2375–2379. doi: 10.1099/0022-1317-76-9-2375. [DOI] [PubMed] [Google Scholar]
  41. Subramanian G., McClain D. S., Perez A., Fuller A. O. Swine testis cells contain functional heparan sulfate but are defective in entry of herpes simplex virus. J Virol. 1994 Sep;68(9):5667–5676. doi: 10.1128/jvi.68.9.5667-5676.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Wittels M., Spear P. G. Penetration of cells by herpes simplex virus does not require a low pH-dependent endocytic pathway. Virus Res. 1991 Mar;18(2-3):271–290. doi: 10.1016/0168-1702(91)90024-p. [DOI] [PubMed] [Google Scholar]
  43. WuDunn D., Spear P. G. Initial interaction of herpes simplex virus with cells is binding to heparan sulfate. J Virol. 1989 Jan;63(1):52–58. doi: 10.1128/jvi.63.1.52-58.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES