Research Paper | Published:

Regulated Expression Allows High Level Production and Secretion of HIV-1 gp120 Envelope Glycoprotein in Drosophila Schneider Cells

Bio/Technologyvolume 9pages173177 (1991) | Download Citation



We have established a stable, continuous culture Drosophila Schneider 2 cell line that efficiently expresses a secreted, truncated form of the HIV envelope gp120 protein in a regulated manner. The Drosophila produced recombinant gp120 protein is highly glycosylated, is recognized by gp120-specific monoclonal antibodies, binds to the CD4 receptor and has the ability to inhibit syncytia formation between uninfected CD4+ cells and HIV infected cells. We conclude that this recombinant Drosophila envelope protein is an appropriate mimic of the authentic viral envelope protein. Thus, the Drosophila cell provides a continuous, stable culture system for the efficient expression of secreted forms of complex surface glycoproteins in quantities sufficient for detailed analyses.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.


  1. 1

    Putney, S.D., Matthews, T.J., Robey, W.G., Lynn, D.L., Robcrt-Guroff, M., Mueller, W.T., Langlois, A.J., Ghrayeb, J., Petteway, S.R., Jr., Weinhold, K.J., Fischinger, P.J., Wong-Staal, F., Gallo, R.C. and Bolognesi, D.P. 1986. HTLV-lII/LAV-neutralizing antibodies to an E. coli a-produced fragment of the virus envelope. Science 244: 1392–1395.

  2. 2

    Chiang, C.S., Grove, T., Cooper, M., Cuan, J., Kowalski, A., Parcells, K., Tsunokawa, M., Rosenberg, M., Arcuri, E., Franklin, S., Smith, T. and Debouck, C.D. 1989. Development of a confirmatory enzyme-linked immunosorbent assay for HIV-1 antibodies. Clin. Chem. 35: 946–952.

  3. 3

    Sammuel, K.P., Seht, A., Zweig, M., Showaker, S.D. and Papas, T.S. 1988. Bacterial expression and characterization of nine polypeptides encoded by segments of the envelope gene of human immunodeficiency virus. Gene 64: 121–134.

  4. 4

    Crowl, R., Ganguly, K., Gordon, M., Conroy, R., Schaber, M., Kramer, R., Shaw, G., Wong-Staal, F. and Reddy, E.P. 1985. HTLV-III env gene products synthesized in E. coli are recognized by antibodies present in the sera of AIDS patients. Cell 41: 979–986.

  5. 5

    Morikawa, Y., Moore, J.P. and Jones, I.M. 1990. HIV-1 envelope protein gp120 expression by secretion in E. coli: assessment of CD4 binding and use in epitope mapping. J. Virol. Methods 29: 105–114.

  6. 6

    Barr, P.D., Steimer, K.S., Sabin, E.A., Parkes, D.M., George-Nascimento, C., Stephens, J.C., Powers, M.A., Gyenes, A., Van Nest, G.A., Miller, E.T., Higgins, K.W. and Luciw, P.A. 1987. Antigenicity and immunogenicity of domains of the human immunodeficiency virus (HIV) envelope polypeptide expressed in the yeast Saccharomyces cerevisiae. Vaccine 5: 90–101.

  7. 7

    Haigwood, N.L., Barker, C.B., Higgins, K.W., Skiles, P.V., Moore, G.K., Mann, K.A., Lee, D.R., Eichberg, J.W. and Steimer, K.S. 1990. Evidence for neutralizing antibodies directed against conformational epitopes of HIV-1 gp120, p. 313–320. In: Vaccines 90. Chanock, R. M., Lerner, R. A., Brown, F. and Ginsbcrg, H. (Eds.). Cold Spring Harbor Press, NY.

  8. 8

    Hu, S.-L., Kosowski, S.G. and Dalrymple, J.M. 1986. Expression of AIDS virus envelope gene in recombinant vaccinia viruses. Nature 320: 537–542.

  9. 9

    Kieny, M.P., Rautmann, S.G., Schmitt, D., Dot, K., Wain-Hobson, S., Alizon, M., Girard, M., Chamaret, S., Laurent, L., Montagnier, L. and Lecocq, J.-P. 1986. AIDS virus env protein expressed from a recombinant vaccinia virus. Bio/Technology 4: 790–795.

  10. 10

    Chakrabarti, S., Robert-Guroff, M., Wong-Staal, F., Gallo, R.C. and Moss, B. 1986. Expression of the HTLV-III envelope gene by a recombinant vaccinia-virus. Nature 320: 535–537.

  11. 11

    Dewar, R.L., Natarajan, V., Vasudevachari, M.B. and Saltzman, N.P. 1989. Synthesis and processing of human deficiency virus type 1 envelope proteins encoded by a recombinant human adenovirus. J. Virol. 63: 129–142.

  12. 12

    Lynn, D.L., Putney, S.D., Carson, H., Matthews, T., Robert-Guroff, M., Krohn, K., Bolognesi, D., Gallo, R. and Rusche, J. 1988. Production of recombinant HIV envelope protein in insect cells and its potential use as a subunit vaccine, p. 359–368. In: Mech. Contr. Gene Exp. Cullen, B. et al. (Eds.). Alan R. Liss, Inc.

  13. 13

    Hu,S.-L., Kosowski, S.G. and Schaff, K. 1987. Expression of envelope glycoproteins of human immunodeficiency virus by an insect virus vector. J. Virol. 61: 3617–3620.

  14. 14

    Wells, D.E. and Compans, R.W. 1990. Expression and characterization of a functional human immunodeficiency virus envelope glyco-protein in insect cells. Virology 176: 575–586.

  15. 15

    Morikawa, Y., Overton, H.A., Moore, J.P., Wilkinson, A.J., Brady, R.L., Lewis, S.J., Jones, I.M. 1990. Expression of HIV-1 gp120 and human soluble CD4 by recombinant baculoviruses and their interaction in vitro. AIDS Res. Human Retrov. 6: 765–773.

  16. 16

    Jarvis, D.L., Fleming, J.G.W., Kovacs, G.R., Summers, M.D. and Guarino, L.A. 1990. Use of early baculovirus promoters for continuous expression and efficient processing of foreign gene products in stably transformed Leipidopteran cells. Bio/Technology 8: 950–955.

  17. 17

    Laskey, L.A., Groopman, J.E., Fennie, C.W., Benz, P.M., Capon, D.J., Dowbenko, D.J., Nakamura, G.R., Nunes, W.M., Renz, M.E. and Berman, P.W. 1986. Neutralization of the AIDS retrovirus by antibodies to a recombinant envelope protein. Science 243: 209–212.

  18. 18

    Johansen, H., van der Straten, A., Sweet, R., Otto, E., Maroni, G. and Rosenberg, M., 1989. Regulated expression at high copy number allows production of a growth-inhibitory oncogene product in Drosophila Schneider cells. Gene and Devel. 3: 882–889.

  19. 19

    Van der Straten, A., Johansen, H., Rosenberg, M. and Sweet, R. 1989. Introduction and constitutive expresion of gene products in cultured Drosophila cells using hygromycin B selection. Curr. Methods Mol. Biol. 1: 1–8.

  20. 20

    Van der Straten, A., Johansen, H., Sweet, R. and Rosenberg, M. 1987. Efficient expression of foreign genes in cultured Drosophila melanogaster cells using hygromycin B selection, p. 131–134. In: Invertebrate Cell Systems Applications. J. Mitsuhashi (Ed.). CRC Press, Boca Raton, FL.

  21. 21

    Conners, R.W., Norverale, J., Sweet, R.W., Berkowitz, B.B., Williams, D., Franklin, S., Shebushki, R., Trill, J.J., Pfarr, D. and Reff, M.E. 1988. Coamplification of tissue plasminogen activator in DHFR positive bivine endothelial cells: in vitro characterization of the purified serine protease. DNA 7: 651–661.

  22. 22

    Opdenakker, G., Weening, H., Colen, D., Billiau, A. and De Somer, P. 1982. Messenger RNA for human tissue plasminogen activator. Eur. J. Biochem. 121: 269–274.

  23. 23

    Muesing, M.A., Smith, D.H., Cabradilla, C.D., Benton, C.V., Las-key, L.A. and Capon, D.J. 1985. Nucleic acid structure and expression of the human AIDS/lymphadenopathy retrovirus. Nature 313: 450–458.

  24. 24

    Allan, J.S., Coligan, J.E., Barin, F., McLane, M.F., Sodroski, J.G., Rosen, C.A., Haseltine, W.A., Lee, T.H., Essex, M. 1985. Major glycoprotein antigens that induce antibodies in AIDS patients are encoded by HTLV-III. Science 228: 1091–1094.

  25. 25

    Veronese, F.M., DeVico, A.L., Copeland, T.D., Oroszlan, S., Gallo, R.C., Sarngadharan, M.G. 1985. Characterization of gp41 as the transmembrane protein coded by the HTLV-III/LAV envelope gene. Science 229: 1402–1405.

  26. 26

    Mizuochi, T., Spellman, M.W., Larkin, M., Solomon, J., Basa, L.J. and Feizi, T. 1988. Carbohydrate structures of the human-immunodeficiency-virus (HIV) recombinant envelope glycoprotein gp120 produced in Chinese-hamster ovary cells. Biochem. J. 254: 599–603.

  27. 27

    Mizuochi, T., Spellman, M.W., Larkin, M., Solomon, J., Basa, L.J. and Feizi, T. 1988. Structural characterization by chromatographic profiling of the oligosaccharides of human immunodeficiency virus (HIV) recombinant envelope glycoprotein gp120 produced in chi-nese-hamster ovary cells. Biomed. Chrom. 2: 260–270.

  28. 28

    Geyer, H., Holschbach, C., Hunsmann, G. and Schneider, J. 1988. Carbohydrates of human immunodeficiency virus: structures of oligosaccharides linked to the envelope glycoprotein 120. J. Biol. Chem. 263: 11760–11767.

  29. 29

    Mizuochi, T., Matthews, T.J., Kato, M., Harnako, J., Titani, K., Solomon, J. and Feizi, T. 1990. Diversity of oligosaccharide structures on the envelope glycoprotein gp120 of human immunodeficiency virus 1 from the lymphoblastoid cell line h9. J. Biol. Chem. 265: 8519–8524.

  30. 30

    Matthews, T.J., Weinhold, K.J., Lyerly, H.K., Langlois, A., Wigzell, H. and Bolognesi, D.P. 1987. Interaction between the human T-cell lymphotropic virus type IIIB envelope glycoprotein gp120 and the surface antigen CD4: Role of carbohydrate in binding and cell fusion. Proc. Natl. Acad. Sci. USA 84: 5424–5428.

  31. 31

    Fennie, C. and Laskey, L.A. 1989. Model for intracellular folding of the human immunodeficiency virus type 1 gp120. J. Virol. 63: 639–646.

  32. 32

    Farrar, W.L., Harel-Bellan, A., Ferris, D.K. 1988. Characterization of CD4 glycoprotein determinant-HIV envelope protein interactions: perspectives for analog and vaccine development. Critical Rev. Immunol. 8: 315–339.

  33. 33

    Fenquillet, E., Clerget-Raslain, B., Gluckman, J.C., Guetard, D., Montagnier, L. and Bahraoui, E. 1989. Role of N-linked glycans in the interactions between the envelope glycoprotein of human immunodeficiency virus and its CD4 cellular receptor. J. Exp. Med. 169: 807–822.

  34. 34

    Fenquillet, E., Gluckman, J.C. and Bahraoui, E. 1990. Role of N-linked glycans of envelope glycoproteins in infectivity of human immunodeficiency virus type 1. J. Virol. 64: 2841–2848.

  35. 35

    Hsieh, P. and Robbins, P.W. 1984. Regulation of asparagine-linked oligosaccharide processing. J. Biol. Chem. 259: 2475–2482.

  36. 36

    Anumula, K.R. and Taylor, P.B. 1989. Rapid N-linked oligosaccharide mapping of glycoproteins using a carbohydrate analyzer. Glyco-con. J. 6: 414.

  37. 37

    Thiriart, C., Francotte, M., Cohen, J., Collognon, C., Delers, A., Kummert, S., Molitor, C., Gilles, D., Roelants, P., van Wijnendaele, F., De Wilde, M. and Bruck, C. 1989. Several antigenic determinants exposed on the gp120 moiety of HIV-1 gp160 are hidden on the mature gp120. J. Immunol. 143: 1832–1842.

  38. 38

    Kowalski, M., Potz, J., Basiripour, L., Dorfman, T., Wei, C.G., Terwillinger, E., Dayton, A., Rosen, C., Haseltine, W. and Sodroski, J. 1987. Functional regions of the envelope glycoproteins of human immunodeficiency virus type 1. Science 247: 1351–1355.

  39. 39

    Mellon, P., Parker, V., Glusman, Y. and Maniatais, T. 1981. Identification of DNA sequences required for transcription of the human globin gene in a new SV40 host-vector system. Cell 27: 279–288.

  40. 40

    Schneider, I. 1972. Cell lines derived from late embryonic stages of Drosophila melanogaster. J. Embryol. Exp. Morphol. 27: 353–356.

  41. 41

    Lindquist, S., Sonoedo, S., Cox, T. and Slusser, K. 1982. Instant medium for Drosophila tissue culture cells. Drosophila Inf. Serv. 58: 163–164.

  42. 42

    Dean, K.C., McDougal, J.S., Inacker, R., Folena-Wasserman, G., Arthos, J., Rosenberg, J., Maddon, P.J., Axel, R., Sweet, R.M. 1988. A soluble form of CD4 (T4) protein inhibits AIDS virus infection. Nature 331: 82–84.

Download references

Author information

Author notes

  1. Jeffery S. Culp: Corresponding author.


  1. Departments of Protein Biochemistry and Gene Expression Sciences, SmithKline Beecham Pharmaceuticals, King of Prussia, Pennsylvania, 19406

    • Jeffery S. Culp
    • , Hanne Johansen
    • , Brian Hellmig
    • , Jeffery Beck
    •  & Martin Rosenberg
  2. Department of Surgery, Duke University Medical Center, Durham, North Carolina, 27710

    • Thomas J. Matthews
  3. Department of Molecular and Cellular Biology, SmithKline Beecham Biologicals, B-1330, Rixensart, Belgium

    • Anne Delers


  1. Search for Jeffery S. Culp in:

  2. Search for Hanne Johansen in:

  3. Search for Brian Hellmig in:

  4. Search for Jeffery Beck in:

  5. Search for Thomas J. Matthews in:

  6. Search for Anne Delers in:

  7. Search for Martin Rosenberg in:

About this article

Publication history



Issue Date


Further reading