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Volume 77, Issue 7

Antigenicity of truncated forms of the human immunodeficiency virus type 1 envelope glycoprotein Free

Abstract

Chinese hamster ovary (CHO) cell lines secreting a series of truncated forms of human immunodeficiency virus type 1 (HIV-1) IIIB (clone BH10) gp120 were assembled. Using purified glyco-proteins, we demonstrated the functional and structural integrity of these truncates by their reactivity with both sCD4 and anti-gp120 monoclonal antibodies (MAbs). Deletion of the V1, V2 and V3 regions had minimal effects on CD4 binding, but deletion of the NH terminus affected the folding of the truncated molecule. Deletion of either V1/V2 or V1/V2/V3 regions led to enhanced recognition by some, but not all, MAbs mapping to the CD4 binding site. In contrast, deletion of the V1/V2 regions had no effect on the ability of V3-specific MAbs to bind to the truncate. These results support the use of truncated forms of gp120 as components of potential HIV vaccines.

  • Received:
  • Accepted:
  • Published Online:
© Journal of General Virology 1996
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1996-07-01
2024-04-25
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References

  1. Bebbington C. R., Renner G., Thomson S., King D., Abrams D., Yarranton G. T. 1992; High-level expression of a recombinant antibody from myeloma cells using a glutamine synthetase gene as an amplifiable selectable marker. Biotechnology 10:169–175
     [Google Scholar]
  2. Brady R. L., Dodson E. J., Dodson G. G., Lange G., Davis S. J., Williams A. F., Barclay A. N. 1993; Crystal structure of domains 3 and 4 of rat CD4: relationship to the NH2 terminal domains. Science 260:979–983
     [Google Scholar]
  3. Carne C., Weller I. V. D., Johnson A. 1987; Prevelance of antibodies to human immunodeficiency virus, gonorrhoea rate, and changes in behaviour in homosexual men in London. Lancet i:6560–6568
     [Google Scholar]
  4. Chapman B. S., Thayer R. M., Vincent K. A., Haigwood N. I. 1991; Effect of intron A from human cytomegalovirus (Towne) intermediate early gene on heterologous expression in mammalian cells. Nucleic Acids Research 19:3979–3986
     [Google Scholar]
  5. Cockett M. I., Bebbington C. R., Yarranton G. T. 1990; High level expression of tissue inhibitor of metalloproteinases in Chinese hamster ovary cells using glutamine synthetase gene amplification. Biotechnology 10:662–667
     [Google Scholar]
  6. Cockett M. I., Bebbington C. R., Yarranton G. T. 1991; The use of engineered EIA genes to transactivate the hCMV-MIE promoter in permanant CHO cell lines. Nucleic Acids Research 19:319–325
     [Google Scholar]
  7. Cordell J., Moore J. P., Dean C. J., Klasse P. J., Weiss R. A., McKeating J. A. 1991; Rat monoclonal antibodies to non-overlapping epitopes of human immunodeficiency virus type 1 gp120 block CD4 binding in vitro. Virology 185:72–79
     [Google Scholar]
  8. Dowbenko D., Nakamura G., Fennie C., Shimasaki C., Riddle L., Harris R., Gregory T., Lasky L. 1988; Epitope mapping of the human immunodeficiency virus type I gp120 with monoclonal antibodies. Journal of Virology 62:4703–4711
     [Google Scholar]
  9. Fenouillet E., Clerget-Raslain B., Gluckman J. C., Guetard D., Montagnier L., Bahraoui E. 1989; Role of N-linked glycans in the interaction between the envelope glycoprotein of human immunodeficiency and its CD4 cellular receptor. Journal of Experimental Medicine 169:807–822
     [Google Scholar]
  10. Kingston R. E., Kaufman R. J., Bebbington C. R., Rolfe M. R. 1992; Amplification using CHO cell expression vectors. In Short Protocols in Molecular Biology16/55–16/62 Edited by Ausubel F. M. and others New York: Wiley;
     [Google Scholar]
  11. Karwowska S., Gorny M. K., Buchbinder A., Gianakakos V., Williams C., Fuerst T., Zolla-Pazner S. 1992; Production of human monoclonal antibodies specific for conformational and linear non-V3 epitopes of gp120. AIDS and Research in Human Retroviruses 8:1099–1106
     [Google Scholar]
  12. Kowalski M., Potz J., Basiripour L., Dorfman T., Goh W. C., Terwilliger E., Dayton A., Rosen C., Haseltine W., Sodroski J. 1987; Functional regions of the envelope glycoprotein of human immunodeficiency virus type 1. Science 237:1351–1355
     [Google Scholar]
  13. Leonard C. K., Spellman M. W., Riddle L., Harris R. J., Thomas J. N., Gregory T. J. 1990; Assignment of intrachain disulphide bonds and characterisation of potential glycosylation sites of the type I recombinant human immunodeficiency virus envelope glycoprotein (gp120) expressed in Chinese hamster ovary cells. Journal of Biological Chemistry 265:10373–10382
     [Google Scholar]
  14. Linsley P. S., Ledbetter J. A., Kinney-Thomas E., Hu S.-L. 1988; Effects of anti-gp120 monoclonal antibodies on CD4 receptor binding by the env protein of human immunodeficiency virus type I. Journal of Virology 62:3695–3702
     [Google Scholar]
  15. McKeating J. A., Thali M., Furman C., Karwowska S., Gorny M. K., Cordell J., Zolla-Pazner S., Sodroski J., Weiss R. A. 1992a; Amino acid residues of the HIV type I gp120 critical for the binding of rat and human neutralizing antibodies that block the gp120 sCD4 interaction. Virology 190:134–142
     [Google Scholar]
  16. McKeating J. A., Cordell J., Dean C. J., Balfe P. 1992b; Synergistic interaction between ligands binding to the CD4 binding site and V3 domain of human immunodeficiency virus type I gp120. Virology 191:732–742
     [Google Scholar]
  17. McKeating J. A., Bennett J., Zolla-Pazner S., Schutten M., Ashelford S., Leigh Brown A., Balfe P. 1993; Resistance of a human serum selected HIV type I escape mutant to neutralization by CD4 binding site monoclonal antibodies is conferred by a single amino acid change in gp120. Journal of Virology 67:5216–5225
     [Google Scholar]
  18. Matsushita S., Robert-Guroff M., Rusche J., Koito A., Sutoh H., Hattori T., Hoshino H., Javaherian K., Jellis C., Takatsuki K., Putney S. 1988; An HIV neutralizing monoclonal antibody and mapping of the neutralizing epitope. Journal of Virology 62:2107–2114
     [Google Scholar]
  19. Matthews T. J., Wild C., Chen C.-H., Bolognesi D. P., Greenberg M. L. 1994; Structural rearrangements in the transmembrane glycoprotein after receptor binding. Immunological Reviews 140:93–104
     [Google Scholar]
  20. Modrow S., Hahn B. W., Shaw G. M., Gallo R. C., Wong-Staal F., Wolf H. 1987; Computer-assisted analysis of envelope protein sequences of seven human immunodeficiency virus isolates: predictions of antigenic epitopes in conserved and variable regions. Journal of Virology 61:570–578
     [Google Scholar]
  21. Moore J. P. 1990; Simple methods for monitoring HIV-1 and HIV-2 gp120 binding to sCD4 by ELISA: HIV-2 has a 25-fold lower affinity than HIV-1 for sCD4. AIDS 4:297–305
     [Google Scholar]
  22. Moore J. P., McKeating J. A., Jones I. M., Stephens P. E., Clements G., Thomson S., Weiss R. A. 1990; Characterisation of recombinant gp120 and gp160 from HIV-1: binding to monoclonal antibodies and sCD4. AIDS 4:307–315
     [Google Scholar]
  23. Moore J. P., Jameson B. A., Sattentau Q. J., Willey R., Sodroski J. 1993; Towards a structure of the HIV1 envelope glycoprotein gp120: an immunochemical approach. Philosophical Transactions of the Royal Society of London, Series B 342:83–88
     [Google Scholar]
  24. Moore J. P., Sattentau Q. J., Wyatt R., Sodroski J. 1994; Probing the structure of the surface glycoprotein gp120 of human immunodeficiency virus type 1 with a panel of monoclonal antibodies. Journal of Virology 68:469–484
     [Google Scholar]
  25. Myers G., Josephs S. F., Rabson A. B., Smith T. F., Wong-Staal F. (editors) 1988; Human Retroviruses and AIDS. Los Alamos, N. Mex: Los Alamos National Laboratory;
     [Google Scholar]
  26. Nakamura G. R., Byrn R., Wilkes D. M., Fox J. A., Hobbs M. R., Hasting R., Wessling H. C., Norcross M. A., Fendly B. M., Berman P. W. 1993; Strain specific and binding affinity requirements of neutralizing monoclonal antibodies to the C4 domain of gp120 from HIV type 1. Journal of Virology 67:6179–6191
     [Google Scholar]
  27. Olshevsky U., Helseth E., Furman C., Li J., Haseltine W., Sodroski J. 1990; Identification of individual human immunodeficiency virus type 1 gp120 amino acids important for CD4 receptor binding. Journal of Virology 64:5701–5707
     [Google Scholar]
  28. Pollard S. R., Meier W., Chow P., Rosa J. J., Wiley D. C. 1991; CD4-binding regions of human immunodeficiency virus envelope glycoprotein gp120 defined by proteolytic digestion. Proceedings of the National Academy of Sciences, USA 88:11320–11324
     [Google Scholar]
  29. Pollard S. R., Rosa M. D., Rosa J. J., Wiley D. C. 1992; Truncated variants of gp120 bind CD4 with high affinity and suggest a minimum CD4 binding region. EMBO Journal 11:585–591
     [Google Scholar]
  30. Ryu S. E., Kwong P. D., Trunen A., Porter T. G., Arthos J., Rosenberg M., Dai X., Xyong N., Axel R., Sweet R. W., Hendrickson W. A. 1990; Crystal structure of an HIV binding recombinant fragment of human CD4. Nature 348:419–426
     [Google Scholar]
  31. Sattentau Q. J., Moore J. P., Vignaux F., Traincard F., Poignard P. 1993; Conformational changes induced in the envelope glycoproteins of human and simian immunodeficiency viruses by soluble receptor binding. Journal of Virology 67:7383–7393
     [Google Scholar]
  32. Starcich B. R., Hahn B. H., Shaw G. M., Mcneely P. D., Modrow S., Wolf H., Parks E. S., Parks W. P., Josephs S. F., Gallo R. C., Wong-Staal F. 1986; Identification and characterisation of conserved and variable regions in the envelope gene of HTLV-III/LAV, the retrovirus of AIDS. Cell 45:637–648
     [Google Scholar]
  33. Syu W. J., Huang J. H., Lee T. H. 1990; The N-terminal region of the human immunodeficiency virus envelope glycoprotein gp120 contains potential binding sites for CD4. Proceedings of the National Academy of Sciences, USA 87:3695–3699
     [Google Scholar]
  34. Thali M., Olshevsky U., Furman C., Gabuzda D., Li J., Sodroski J. 1991a; Effects of changes in gp120-CD4 binding affinity on human immunodeficiency virus type 1 envelope glycoprotein function and soluble CD4 sensitivity. Journal of Virology 65:5007–5012
     [Google Scholar]
  35. Thali M., Olshevsky U., Furman C., Gabuzda D., Posner M., Sodroski J. 1991b; Characterization of a discontinuous human immunodeficiency virus type 1 gp120 epitope recognized by a broadly neutralizing human monoclonal antibody. Journal of Virology 65:6188–6193
     [Google Scholar]
  36. Thali M., Furman C., Ho D. D., Robinson J., Tilley S., Pinter A., Sodroski J. 1992; Discontinuous, conserved neutralization epitopes overlapping the CD4 binding region of the human immunodeficiency virus type I gp120 envelope glycoprotein. Journal of Virology 66:5635–5641
     [Google Scholar]
  37. Thali M., Moore J. P., Furman C., MacArthur C., Ho D. D., Robinson J., Sodroski J. 1993; Characterization of conserved human immunodeficiency virus type 1 gp120 neutralization epitopes exposed upon gp120-CD4 binding. Journal of Virology 67:3978–3988
     [Google Scholar]
  38. Wang J., Yan Y., Garrett R. J., Liu J., Rodgers D. W., Garlick R. L., Tarr G. E., Husain Y., Reinherz E. L., Harrison S. C. 1990; Atomic structure of a fragment of human CD4 containing two immunoglobulin like domains. Nature 348:411–418
     [Google Scholar]
  39. Wyatt R., Sullivan N., Thali M., Repke H., Ho D., Robinson J., Posner M., Sodroski J. 1993; Functional and immunologic characterization of human immunodeficiency virus type 1 envelope glycoproteins containing deletions of the major variable regions. Journal of Virology 67:4557–4565
     [Google Scholar]
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Antigenicity of truncated forms of the human immunodeficiency virus type 1 envelope glycoprotein
J. Gen. Virol. 77, 1403 (1996); https://doi.org/10.1099/0022-1317-77-7-1403
/content/journal/jgv/10.1099/0022-1317-77-7-1403
/content/journal/jgv/10.1099/0022-1317-77-7-1403
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