Skip to main content
PMC home page
Immunology logoLink to Immunology
. 1992 Jul;76(3):367–372.

Constitutively expressing cell lines that secrete a truncated bovine herpes virus-1 glycoprotein (gpI) stimulate T-lymphocyte responsiveness.

T P Leary 1, Y Gao 1, G A Splitter 1
PMCID: PMC1421700  PMID: 1526647

Abstract

The desire to obtain authentically glycosylated viral protein products in sufficient quantity for immunological study has led to the use of eucaryotic expression vectors for protein production. An additional advantage is that these protein products can be studied individually in the absence of their native viral environment. We have cloned a complementary DNA (cDNA) encoding bovine herpes virus-1 (BHV-1) glycoprotein 1 (gpI) into the eucaryotic expression vector, pZipNeo SVX1. Since this protein is normally embedded within the membrane of BHV-1 infected cells, we removed sequences encoding the transmembrane domain of the native protein. After transfection of the plasmid construct into the canine osteosarcoma cell line, D17, or Madin-Darby bovine kidney (MDBK) cells, a truncated BHV-1 (gpI) was secreted into the culture medium as demonstrated by radioimmunoprecipitation and SDS-PAGE. Both a CD4+ T-lymphocyte line specific for BHV-1 and freshly isolated T lymphocytes could recognize and respond to the secreted recombinant gpI. Further, recombinant gpI could elicit both antibody and cellular responses in cattle when used as an immunogen. Having established constitutively glycoprotein producing cell lines, future studies in vaccine evaluation of gpI will be facilitated.

Full text

PDF
367

Images in this article

368Figure 1
on p.368
369Figure 2
on p.369

Selected References

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

  1. Alt F. W., Bothwell A. L., Knapp M., Siden E., Mather E., Koshland M., Baltimore D. Synthesis of secreted and membrane-bound immunoglobulin mu heavy chains is directed by mRNAs that differ at their 3' ends. Cell. 1980 Jun;20(2):293–301. doi: 10.1016/0092-8674(80)90615-7. [DOI] [PubMed] [Google Scholar]
  2. Cantin E. M., Eberle R., Baldick J. L., Moss B., Willey D. E., Notkins A. L., Openshaw H. Expression of herpes simplex virus 1 glycoprotein B by a recombinant vaccinia virus and protection of mice against lethal herpes simplex virus 1 infection. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5908–5912. doi: 10.1073/pnas.84.16.5908. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cepko C. L., Roberts B. E., Mulligan R. C. Construction and applications of a highly transmissible murine retrovirus shuttle vector. Cell. 1984 Jul;37(3):1053–1062. doi: 10.1016/0092-8674(84)90440-9. [DOI] [PubMed] [Google Scholar]
  4. Chan W. L., Lukig M. L., Liew F. Y. Helper T cells induced by an immunopurified herpes simplex virus type I (HSV-I) 115 kilodalton glycoprotein (gB) protect mice against HSV-I infection. J Exp Med. 1985 Oct 1;162(4):1304–1318. doi: 10.1084/jem.162.4.1304. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chase C. C., Carter-Allen K., Letchworth G. J., 3rd The effect of bovine herpesvirus type 1 glycoproteins gI and gIII on herpesvirus infections. J Gen Virol. 1989 Jun;70(Pt 6):1561–1569. doi: 10.1099/0022-1317-70-6-1561. [DOI] [PubMed] [Google Scholar]
  6. Early P., Rogers J., Davis M., Calame K., Bond M., Wall R., Hood L. Two mRNAs can be produced from a single immunoglobulin mu gene by alternative RNA processing pathways. Cell. 1980 Jun;20(2):313–319. doi: 10.1016/0092-8674(80)90617-0. [DOI] [PubMed] [Google Scholar]
  7. Gething M. J., Sambrook J. Construction of influenza haemagglutinin genes that code for intracellular and secreted forms of the protein. Nature. 1982 Dec 16;300(5893):598–603. doi: 10.1038/300598a0. [DOI] [PubMed] [Google Scholar]
  8. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  9. Hoffmann J. W., Steffen D., Gusella J., Tabin C., Bird S., Cowing D., Weinberg R. A. DNA methylation affecting the expression of murine leukemia proviruses. J Virol. 1982 Oct;44(1):144–157. doi: 10.1128/jvi.44.1.144-157.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Israel B. A., Herber R., Gao Y., Letchworth G. J., 3rd Induction of a mucosal barrier to bovine herpesvirus 1 replication in cattle. Virology. 1992 May;188(1):256–264. doi: 10.1016/0042-6822(92)90755-e. [DOI] [PubMed] [Google Scholar]
  11. Israel B. A., Marshall R. L., Letchworth G. J., 3rd Epitope specificity and protective efficacy of the bovine immune response to bovine herpesvirus-1 glycoprotein vaccines. Vaccine. 1988 Aug;6(4):349–356. doi: 10.1016/0264-410x(88)90182-x. [DOI] [PubMed] [Google Scholar]
  12. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  13. Leary T. P., Splitter G. A. Recombinant herpesviral proteins produced by cell-free translation provide a novel approach for the mapping of T lymphocyte epitopes. J Immunol. 1990 Jul 15;145(2):718–723. [PubMed] [Google Scholar]
  14. Liang X. P., Lamm M. E., Nedrud J. G. Oral administration of cholera toxin-Sendai virus conjugate potentiates gut and respiratory immunity against Sendai virus. J Immunol. 1988 Sep 1;141(5):1495–1501. [PubMed] [Google Scholar]
  15. Marshall R. L., Israel B. A., Letchworth G. J., 3rd Monoclonal antibody analysis of bovine herpesvirus-1 glycoprotein antigenic areas relevant to natural infection. Virology. 1988 Aug;165(2):338–347. doi: 10.1016/0042-6822(88)90578-8. [DOI] [PubMed] [Google Scholar]
  16. Marshall R. L., Rodriguez L. L., Letchworth G. J., 3rd Characterization of envelope proteins of infectious bovine rhinotracheitis virus (bovine herpesvirus 1) by biochemical and immunological methods. J Virol. 1986 Mar;57(3):745–753. doi: 10.1128/jvi.57.3.745-753.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Martin S., Moss B., Berman P. W., Laskey L. A., Rouse B. T. Mechanisms of antiviral immunity induced by a vaccinia virus recombinant expressing herpes simplex virus type 1 glycoprotein D: cytotoxic T cells. J Virol. 1987 Mar;61(3):726–734. doi: 10.1128/jvi.61.3.726-734.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Misra V., Gilchrist J. E., Weinmaster G., Qualtiere L., Van den Hurk S., Babiuk L. A. Herpesvirus-induced "early" glycoprotein: characterization and possible role in immune cytolysis. J Virol. 1982 Sep;43(3):1046–1054. doi: 10.1128/jvi.43.3.1046-1054.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Misra V., Nelson R., Smith M. Sequence of a bovine herpesvirus type-1 glycoprotein gene that is homologous to the herpes simplex gene for the glycoprotein gB. Virology. 1988 Oct;166(2):542–549. doi: 10.1016/0042-6822(88)90525-9. [DOI] [PubMed] [Google Scholar]
  20. Murphy A. J., Efstratiadis A. Cloning vectors for expression of cDNA libraries in mammalian cells. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8277–8281. doi: 10.1073/pnas.84.23.8277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Nedrud J. G., Liang X. P., Hague N., Lamm M. E. Combined oral/nasal immunization protects mice from Sendai virus infection. J Immunol. 1987 Nov 15;139(10):3484–3492. [PubMed] [Google Scholar]
  22. Pachl C., Burke R. L., Stuve L. L., Sanchez-Pescador L., Van Nest G., Masiarz F., Dina D. Expression of cell-associated and secreted forms of herpes simplex virus type 1 glycoprotein gB in mammalian cells. J Virol. 1987 Feb;61(2):315–325. doi: 10.1128/jvi.61.2.315-325.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Palmer L. D., Leary T. P., Wilson D. M., Splitter G. A. Bovine natural killer-like cell responses against cell lines expressing recombinant bovine herpesvirus type 1 glycoproteins. J Immunol. 1990 Aug 1;145(3):1009–1014. [PubMed] [Google Scholar]
  24. Rose J. K., Bergmann J. E. Expression from cloned cDNA of cell-surface secreted forms of the glycoprotein of vesicular stomatitis virus in eucaryotic cells. Cell. 1982 Oct;30(3):753–762. doi: 10.1016/0092-8674(82)90280-x. [DOI] [PubMed] [Google Scholar]
  25. Rosenthal K. L., Smiley J. R., South S., Johnson D. C. Cells expressing herpes simplex virus glycoprotein gC but not gB, gD, or gE are recognized by murine virus-specific cytotoxic T lymphocytes. J Virol. 1987 Aug;61(8):2438–2447. doi: 10.1128/jvi.61.8.2438-2447.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sethi K. K., Omata Y., Schneweis K. E. Protection of mice from fatal herpes simplex virus type 1 infection by adoptive transfer of cloned virus-specific and H-2-restricted cytotoxic T lymphocytes. J Gen Virol. 1983 Feb;64(Pt 2):443–447. doi: 10.1099/0022-1317-64-2-443. [DOI] [PubMed] [Google Scholar]
  27. Splitter G. A., Eskra L., Abruzzini A. F. Cloned bovine cytolytic T cells recognize bovine herpes virus-1 in a genetically restricted, antigen-specific manner. Immunology. 1988 Jan;63(1):145–150. [PMC free article] [PubMed] [Google Scholar]
  28. Tikoo S. K., Fitzpatrick D. R., Babiuk L. A., Zamb T. J. Molecular cloning, sequencing, and expression of functional bovine herpesvirus 1 glycoprotein gIV in transfected bovine cells. J Virol. 1990 Oct;64(10):5132–5142. doi: 10.1128/jvi.64.10.5132-5142.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Whitbeck J. C., Bello L. J., Lawrence W. C. Comparison of the bovine herpesvirus 1 gI gene and the herpes simplex virus type 1 gB gene. J Virol. 1988 Sep;62(9):3319–3327. doi: 10.1128/jvi.62.9.3319-3327.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Yates W. D. A review of infectious bovine rhinotracheitis, shipping fever pneumonia and viral-bacterial synergism in respiratory disease of cattle. Can J Comp Med. 1982 Jul;46(3):225–263. [PMC free article] [PubMed] [Google Scholar]
  31. Zarling J. M., Moran P. A., Burke R. L., Pachl C., Berman P. W., Lasky L. A. Human cytotoxic T cell clones directed against herpes simplex virus-infected cells. IV. Recognition and activation by cloned glycoproteins gB and gD. J Immunol. 1986 Jun 15;136(12):4669–4673. [PubMed] [Google Scholar]
  32. Zarling J. M., Moran P. A., Lasky L. A., Moss B. Herpes simplex virus (HSV)-specific human T-cell clones recognize HSV glycoprotein D expressed by a recombinant vaccinia virus. J Virol. 1986 Aug;59(2):506–509. doi: 10.1128/jvi.59.2.506-509.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. van Drunen Littel-van den Hurk S., Gifford G. A., Babiuk L. A. Epitope specificity of the protective immune response induced by individual bovine herpesvirus-1 glycoproteins. Vaccine. 1990 Aug;8(4):358–368. doi: 10.1016/0264-410x(90)90095-4. [DOI] [PubMed] [Google Scholar]
  34. van Drunen Littel-van den Hurk S., van den Hurk J. V., Babiuk L. A. Topographical analysis of bovine herpesvirus type-1 glycoproteins: use of monoclonal antibodies to identify and characterize functional epitopes. Virology. 1985 Jul 15;144(1):216–227. doi: 10.1016/0042-6822(85)90319-8. [DOI] [PubMed] [Google Scholar]

Articles from Immunology are provided here courtesy of British Society for Immunology

RESOURCES