Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1987 Jun;61(6):1796–1807. doi: 10.1128/jvi.61.6.1796-1807.1987

Expression of the Epstein-Barr virus gp350/220 gene in rodent and primate cells.

Y Whang, M Silberklang, A Morgan, S Munshi, A B Lenny, R W Ellis, E Kieff
PMCID: PMC254182  PMID: 3033311

Abstract

The gene encoding the Epstein-Barr virus envelope glycoproteins gp350 and gp220 was inserted downstream of the cytomegalovirus immediate-early, Moloney murine leukemia virus, mouse mammary tumor virus, or varicella-zoster virus gpI promoters in vectors containing selectable markers. Host cell and recombinant vector systems were defined which enabled the isolation of rodent or primate cell clones which expressed gp350/220 in substantial quantities. Continued expression of gp350/220 required maintenance of cells under positive selection for linked markers and periodic cloning. gp350/220 expressed in various host cells varied slightly in electrophoretic mobility, probably reflecting differences in glycosylation. Insertion of a stop codon into the gp350/220 open reading frame, upstream of the putative membrane anchor sequence, resulted in efficient secretion of truncated gp350 and gp220 from rat pituitary (GH3) cells. gp350/220 expressed in mammalian cells is highly immunogenic and elicits virus-neutralizing antibodies when administered to mice.

Full text

PDF
1796

Images in this article

Selected References

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

  1. Beisel C., Tanner J., Matsuo T., Thorley-Lawson D., Kezdy F., Kieff E. Two major outer envelope glycoproteins of Epstein-Barr virus are encoded by the same gene. J Virol. 1985 Jun;54(3):665–674. doi: 10.1128/jvi.54.3.665-674.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Biggin M., Farrell P. J., Barrell B. G. Transcription and DNA sequence of the BamHI L fragment of B95-8 Epstein-Barr virus. EMBO J. 1984 May;3(5):1083–1090. doi: 10.1002/j.1460-2075.1984.tb01933.x. [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. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  5. Chou P. Y., Fasman G. D. Empirical predictions of protein conformation. Annu Rev Biochem. 1978;47:251–276. doi: 10.1146/annurev.bi.47.070178.001343. [DOI] [PubMed] [Google Scholar]
  6. Colbère-Garapin F., Horodniceanu F., Kourilsky P., Garapin A. C. A new dominant hybrid selective marker for higher eukaryotic cells. J Mol Biol. 1981 Jul 25;150(1):1–14. doi: 10.1016/0022-2836(81)90321-1. [DOI] [PubMed] [Google Scholar]
  7. Coope D., Heston L., Brandsma J., Miller G. Cross-neutralization of infectious mononucleosis and Burkitt lymphoma strains of Epstein-Barr virus with hyperimmune rabbit antisera. J Immunol. 1979 Jul;123(1):232–238. [PubMed] [Google Scholar]
  8. Dambaugh T., Beisel C., Hummel M., King W., Fennewald S., Cheung A., Heller M., Raab-Traub N., Kieff E. Epstein-Barr virus (B95-8) DNA VII: molecular cloning and detailed mapping. Proc Natl Acad Sci U S A. 1980 May;77(5):2999–3003. doi: 10.1073/pnas.77.5.2999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Davison A. J. DNA sequence of the US component of the varicella-zoster virus genome. EMBO J. 1983;2(12):2203–2209. doi: 10.1002/j.1460-2075.1983.tb01724.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. DiMaio D., Treisman R., Maniatis T. Bovine papillomavirus vector that propagates as a plasmid in both mouse and bacterial cells. Proc Natl Acad Sci U S A. 1982 Jul;79(13):4030–4034. doi: 10.1073/pnas.79.13.4030. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dolyniuk M., Pritchett R., Kieff E. Proteins of Epstein-Barr virus. I. Analysis of the polypeptides of purified enveloped Epstein-Barr virus. J Virol. 1976 Mar;17(3):935–949. doi: 10.1128/jvi.17.3.935-949.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dolyniuk M., Wolff E., Kieff E. Proteins of Epstein-Barr Virus. II. Electrophoretic analysis of the polypeptides of the nucleocapsid and the glucosamine- and polysaccharide-containing components of enveloped virus. J Virol. 1976 Apr;18(1):289–297. doi: 10.1128/jvi.18.1.289-297.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Edson C. M., Thorley-Lawson D. A. Epstein-Barr virus membrane antigens: characterization, distribution, and strain differences. J Virol. 1981 Jul;39(1):172–184. doi: 10.1128/jvi.39.1.172-184.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Edson C. M., Thorley-Lawson D. A. Synthesis and processing of the three major envelope glycoproteins of Epstein-Barr virus. J Virol. 1983 May;46(2):547–556. doi: 10.1128/jvi.46.2.547-556.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ellis R. W., Keller P. M., Lowe R. S., Zivin R. A. Use of a bacterial expression vector to map the varicella-zoster virus major glycoprotein gene, gC. J Virol. 1985 Jan;53(1):81–88. doi: 10.1128/jvi.53.1.81-88.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Epstein M. A., Morgan A. J., Finerty S., Randle B. J., Kirkwood J. K. Protection of cottontop tamarins against Epstein-Barr virus-induced malignant lymphoma by a prototype subunit vaccine. Nature. 1985 Nov 21;318(6043):287–289. doi: 10.1038/318287a0. [DOI] [PubMed] [Google Scholar]
  17. Epstein M. A., Randle B. J., Finerty S., Kirkwood J. K. Not all potently neutralizing, vaccine-induced antibodies to Epstein-Barr virus ensure protection of susceptible experimental animals. Clin Exp Immunol. 1986 Mar;63(3):485–490. [PMC free article] [PubMed] [Google Scholar]
  18. Fingeroth J. D., Weis J. J., Tedder T. F., Strominger J. L., Biro P. A., Fearon D. T. Epstein-Barr virus receptor of human B lymphocytes is the C3d receptor CR2. Proc Natl Acad Sci U S A. 1984 Jul;81(14):4510–4514. doi: 10.1073/pnas.81.14.4510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Harris M. Induction of thymidine kinase in enzyme-deficient Chinese hamster cells. Cell. 1982 Jun;29(2):483–492. doi: 10.1016/0092-8674(82)90165-9. [DOI] [PubMed] [Google Scholar]
  20. Heller M., Dambaugh T., Kieff E. Epstein-Barr virus DNA. IX. Variation among viral DNAs from producer and nonproducer infected cells. J Virol. 1981 May;38(2):632–648. doi: 10.1128/jvi.38.2.632-648.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hoffman G. J., Lazarowitz S. G., Hayward S. D. Monoclonal antibody against a 250,000-dalton glycoprotein of Epstein-Barr virus identifies a membrane antigen and a neutralizing antigen. Proc Natl Acad Sci U S A. 1980 May;77(5):2979–2983. doi: 10.1073/pnas.77.5.2979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hummel M., Thorley-Lawson D., Kieff E. An Epstein-Barr virus DNA fragment encodes messages for the two major envelope glycoproteins (gp350/300 and gp220/200). J Virol. 1984 Feb;49(2):413–417. doi: 10.1128/jvi.49.2.413-417.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Karin M., Haslinger A., Holtgreve H., Richards R. I., Krauter P., Westphal H. M., Beato M. Characterization of DNA sequences through which cadmium and glucocorticoid hormones induce human metallothionein-IIA gene. Nature. 1984 Apr 5;308(5959):513–519. doi: 10.1038/308513a0. [DOI] [PubMed] [Google Scholar]
  24. Kornfeld R., Kornfeld S. Assembly of asparagine-linked oligosaccharides. Annu Rev Biochem. 1985;54:631–664. doi: 10.1146/annurev.bi.54.070185.003215. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Lee F., Mulligan R., Berg P., Ringold G. Glucocorticoids regulate expression of dihydrofolate reductase cDNA in mouse mammary tumour virus chimaeric plasmids. Nature. 1981 Nov 19;294(5838):228–232. doi: 10.1038/294228a0. [DOI] [PubMed] [Google Scholar]
  27. Littman D. R., Thomas Y., Maddon P. J., Chess L., Axel R. The isolation and sequence of the gene encoding T8: a molecule defining functional classes of T lymphocytes. Cell. 1985 Feb;40(2):237–246. doi: 10.1016/0092-8674(85)90138-2. [DOI] [PubMed] [Google Scholar]
  28. Lopata M. A., Cleveland D. W., Sollner-Webb B. High level transient expression of a chloramphenicol acetyl transferase gene by DEAE-dextran mediated DNA transfection coupled with a dimethyl sulfoxide or glycerol shock treatment. Nucleic Acids Res. 1984 Jul 25;12(14):5707–5717. doi: 10.1093/nar/12.14.5707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Mackett M., Arrand J. R. Recombinant vaccinia virus induces neutralising antibodies in rabbits against Epstein-Barr virus membrane antigen gp340. EMBO J. 1985 Dec 1;4(12):3229–3234. doi: 10.1002/j.1460-2075.1985.tb04070.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Meckstroth K. L., Reiss E., Keller J. W., Kaufman L. Detection of antibodies and antigenemia in leukemic patients with candidiasis by enzyme-linked immunosorbent assay. J Infect Dis. 1981 Jul;144(1):24–32. doi: 10.1093/infdis/144.1.24. [DOI] [PubMed] [Google Scholar]
  31. Meuller-Lantzsch N., Georg B., Yamamoto N., zur Hausen H. Epstein-Barr virus-induced proteins. II. Analysis of surface polypeptides from EBV-producing and -superinfected cells by immunoprecipitation. Virology. 1980 Apr 30;102(2):401–411. doi: 10.1016/0042-6822(80)90107-5. [DOI] [PubMed] [Google Scholar]
  32. Montagnon B. J., Fanget B., Nicolas A. J. The large-scale cultivation of VERO cells in micro-carrier culture for virus vaccine production. Preliminary results for killed poliovirus vaccine. Dev Biol Stand. 1981;47:55–64. [PubMed] [Google Scholar]
  33. Morgan A. J., North J. R., Epstein M. A. Purification and properties of the gp340 component of Epstein-Barr virus membrane antigen in an immunogenic form. J Gen Virol. 1983 Feb;64(Pt 2):455–460. doi: 10.1099/0022-1317-64-2-455. [DOI] [PubMed] [Google Scholar]
  34. Morgan A. J., Smith A. R., Barker R. N., Epstein M. A. A structural investigation of the Epstein-Barr (EB) virus membrane antigen glycoprotein, gp340. J Gen Virol. 1984 Feb;65(Pt 2):397–404. doi: 10.1099/0022-1317-65-2-397. [DOI] [PubMed] [Google Scholar]
  35. Mulligan R. C., Berg P. Selection for animal cells that express the Escherichia coli gene coding for xanthine-guanine phosphoribosyltransferase. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2072–2076. doi: 10.1073/pnas.78.4.2072. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Nemerow G. R., Wolfert R., McNaughton M. E., Cooper N. R. Identification and characterization of the Epstein-Barr virus receptor on human B lymphocytes and its relationship to the C3d complement receptor (CR2). J Virol. 1985 Aug;55(2):347–351. doi: 10.1128/jvi.55.2.347-351.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. North J. R., Morgan A. J., Epstein M. A. Observations on the EB virus envelope and virus-determined membrane antigen (MA) polypeptides. Int J Cancer. 1980 Aug;26(2):231–240. doi: 10.1002/ijc.2910260216. [DOI] [PubMed] [Google Scholar]
  38. Pasleau F., Tocci M. J., Leung F., Kopchick J. J. Growth hormone gene expression in eukaryotic cells directed by the Rous sarcoma virus long terminal repeat or cytomegalovirus immediate-early promoter. Gene. 1985;38(1-3):227–232. doi: 10.1016/0378-1119(85)90221-5. [DOI] [PubMed] [Google Scholar]
  39. Qualtiere L. F., Chase R., Pearson G. R. Purification and biologic characterization of a major Epstein Barr virus-induced membrane glycoprotein. J Immunol. 1982 Aug;129(2):814–818. [PubMed] [Google Scholar]
  40. Qualtiere L. F., Pearson G. R. Radioimmune precipitation study comparing the Epstein-Barr virus membrane antigens expressed on P3HR-1 virus-superinfected Raji cells to those expressed on cells in a B-95 virus-transformed producer culture activated with tumor-promoting agent (TPA). Virology. 1980 Apr 30;102(2):360–369. doi: 10.1016/0042-6822(80)90103-8. [DOI] [PubMed] [Google Scholar]
  41. Sanger F., Coulson A. R., Barrell B. G., Smith A. J., Roe B. A. Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol. 1980 Oct 25;143(2):161–178. doi: 10.1016/0022-2836(80)90196-5. [DOI] [PubMed] [Google Scholar]
  42. Shinnick T. M., Lerner R. A., Sutcliffe J. G. Nucleotide sequence of Moloney murine leukaemia virus. Nature. 1981 Oct 15;293(5833):543–548. doi: 10.1038/293543a0. [DOI] [PubMed] [Google Scholar]
  43. Strnad B. C., Adams M. R., Rabin H. Glycosylation pathways of two major Epstein-Barr virus membrane antigens. Virology. 1983 May;127(1):168–176. doi: 10.1016/0042-6822(83)90381-1. [DOI] [PubMed] [Google Scholar]
  44. Strnad B. C., Neubauer R. H., Rabin H., Mazur R. A. Correlation between Epstein-Barr virus membrane antigen and three large cell surface glycoproteins. J Virol. 1979 Dec;32(3):885–894. doi: 10.1128/jvi.32.3.885-894.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Strnad B. C., Schuster T., Klein R., Hopkins R. F., 3rd, Witmer T., Neubauer R. H., Rabin H. Production and characterization of monoclonal antibodies against the Epstein-Barr virus membrane antigen. J Virol. 1982 Jan;41(1):258–264. doi: 10.1128/jvi.41.1.258-264.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Subramani S., Mulligan R., Berg P. Expression of the mouse dihydrofolate reductase complementary deoxyribonucleic acid in simian virus 40 vectors. Mol Cell Biol. 1981 Sep;1(9):854–864. doi: 10.1128/mcb.1.9.854. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Tashjian A. H., Jr, Yasumura Y., Levine L., Sato G. H., Parker M. L. Establishment of clonal strains of rat pituitary tumor cells that secrete growth hormone. Endocrinology. 1968 Feb;82(2):342–352. doi: 10.1210/endo-82-2-342. [DOI] [PubMed] [Google Scholar]
  48. Thomsen D. R., Stenberg R. M., Goins W. F., Stinski M. F. Promoter-regulatory region of the major immediate early gene of human cytomegalovirus. Proc Natl Acad Sci U S A. 1984 Feb;81(3):659–663. doi: 10.1073/pnas.81.3.659. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Thorley-Lawson D. A. Characterization of cross-reacting antigens on the Epstein-Barr virus envelope and plasma membranes of producer cells. Cell. 1979 Jan;16(1):33–42. doi: 10.1016/0092-8674(79)90185-5. [DOI] [PubMed] [Google Scholar]
  50. Thorley-Lawson D. A., Geilinger K. Monoclonal antibodies against the major glycoprotein (gp350/220) of Epstein-Barr virus neutralize infectivity. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5307–5311. doi: 10.1073/pnas.77.9.5307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Thorley-Lawson D. A., Poodry C. A. Identification and isolation of the main component (gp350-gp220) of Epstein-Barr virus responsible for generating neutralizing antibodies in vivo. J Virol. 1982 Aug;43(2):730–736. doi: 10.1128/jvi.43.2.730-736.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Urlaub G., Chasin L. A. Isolation of Chinese hamster cell mutants deficient in dihydrofolate reductase activity. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4216–4220. doi: 10.1073/pnas.77.7.4216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Wells A., Koide N., Klein G. Two large virion envelope glycoproteins mediate Epstein-Barr virus binding to receptor-positive cells. J Virol. 1982 Jan;41(1):286–297. doi: 10.1128/jvi.41.1.286-297.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Wigler M., Pellicer A., Silverstein S., Axel R., Urlaub G., Chasin L. DNA-mediated transfer of the adenine phosphoribosyltransferase locus into mammalian cells. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1373–1376. doi: 10.1073/pnas.76.3.1373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Woychik R. P., Camper S. A., Lyons R. H., Horowitz S., Goodwin E. C., Rottman F. M. Cloning and nucleotide sequencing of the bovine growth hormone gene. Nucleic Acids Res. 1982 Nov 25;10(22):7197–7210. doi: 10.1093/nar/10.22.7197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. de-Thé G., Geser A., Day N. E., Tukei P. M., Williams E. H., Beri D. P., Smith P. G., Dean A. G., Bronkamm G. W., Feorino P. Epidemiological evidence for causal relationship between Epstein-Barr virus and Burkitt's lymphoma from Ugandan prospective study. Nature. 1978 Aug 24;274(5673):756–761. doi: 10.1038/274756a0. [DOI] [PubMed] [Google Scholar]
  57. van Santen V., Cheung A., Kieff E. Epstein-Barr virus RNA VII: size and direction of transcription of virus-specified cytoplasmic RNAs in a transformed cell line. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1930–1934. doi: 10.1073/pnas.78.3.1930. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. von Heijne G. Patterns of amino acids near signal-sequence cleavage sites. Eur J Biochem. 1983 Jun 1;133(1):17–21. doi: 10.1111/j.1432-1033.1983.tb07424.x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES