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An engineered Saccharomyces cerevisiae strain binds the broadly neutralizing human immunodeficiency virus type 1 antibody 2G12 and elicits mannose-specific gp120-binding antibodies.

Publication ,  Journal Article
Luallen, RJ; Lin, J; Fu, H; Cai, KK; Agrawal, C; Mboudjeka, I; Lee, F-H; Montefiori, D; Smith, DF; Doms, RW; Geng, Y
Published in: J Virol
July 2008

The glycan shield of the human immunodeficiency virus type 1 (HIV-1) envelope (Env) protein serves as a barrier to antibody-mediated neutralization and plays a critical role in transmission and infection. One of the few broadly neutralizing HIV-1 antibodies, 2G12, binds to a carbohydrate epitope consisting of an array of high-mannose glycans exposed on the surface of the gp120 subunit of the Env protein. To produce proteins with exclusively high-mannose carbohydrates, we generated a mutant strain of Saccharomyces cerevisiae by deleting three genes in the N-glycosylation pathway, Och1, Mnn1, and Mnn4. Glycan profiling revealed that N-glycans produced by this mutant were almost exclusively Man(8)GlcNAc(2), and four endogenous glycoproteins that were efficiently recognized by the 2G12 antibody were identified. These yeast proteins, like HIV-1 gp120, contain a large number and high density of N-linked glycans, with glycosidase digestion abrogating 2G12 cross-reactivity. Immunization of rabbits with whole Delta och1 Delta mnn1 Delta mnn4 yeast cells produced sera that recognized a broad range of HIV-1 and simian immunodeficiency virus (SIV) Env glycoproteins, despite no HIV/SIV-related proteins being used in the immunization procedure. Analyses of one of these sera on a glycan array showed strong binding to glycans with terminal Man alpha1,2Man residues, and binding to gp120 was abrogated by glycosidase removal of high-mannose glycans and terminal Man alpha1,2Man residues, similar to 2G12. Since S. cerevisiae is genetically pliable and can be grown easily and inexpensively, it will be possible to produce new immunogens that recapitulate the 2G12 epitope and may make the glycan shield of HIV Env a practical target for vaccine development.

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Published In

J Virol

DOI

EISSN

1098-5514

Publication Date

July 2008

Volume

82

Issue

13

Start / End Page

6447 / 6457

Location

United States

Related Subject Headings

  • Virology
  • Saccharomyces cerevisiae
  • Molecular Sequence Data
  • Microscopy, Fluorescence
  • Mannose
  • Immunoprecipitation
  • Immune Sera
  • HIV-1
  • HIV Envelope Protein gp120
  • HIV Antibodies
 

Citation

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Luallen, R. J., Lin, J., Fu, H., Cai, K. K., Agrawal, C., Mboudjeka, I., … Geng, Y. (2008). An engineered Saccharomyces cerevisiae strain binds the broadly neutralizing human immunodeficiency virus type 1 antibody 2G12 and elicits mannose-specific gp120-binding antibodies. J Virol, 82(13), 6447–6457. https://doi.org/10.1128/JVI.00412-08
Luallen, Robert J., Jianqiao Lin, Hu Fu, Karen K. Cai, Caroline Agrawal, Innocent Mboudjeka, Fang-Hua Lee, et al. “An engineered Saccharomyces cerevisiae strain binds the broadly neutralizing human immunodeficiency virus type 1 antibody 2G12 and elicits mannose-specific gp120-binding antibodies.J Virol 82, no. 13 (July 2008): 6447–57. https://doi.org/10.1128/JVI.00412-08.
Luallen RJ, Lin J, Fu H, Cai KK, Agrawal C, Mboudjeka I, Lee F-H, Montefiori D, Smith DF, Doms RW, Geng Y. An engineered Saccharomyces cerevisiae strain binds the broadly neutralizing human immunodeficiency virus type 1 antibody 2G12 and elicits mannose-specific gp120-binding antibodies. J Virol. 2008 Jul;82(13):6447–6457.

Published In

J Virol

DOI

EISSN

1098-5514

Publication Date

July 2008

Volume

82

Issue

13

Start / End Page

6447 / 6457

Location

United States

Related Subject Headings

  • Virology
  • Saccharomyces cerevisiae
  • Molecular Sequence Data
  • Microscopy, Fluorescence
  • Mannose
  • Immunoprecipitation
  • Immune Sera
  • HIV-1
  • HIV Envelope Protein gp120
  • HIV Antibodies