Site-Specific Glycosylation of Virion-Derived HIV-1 Env Is Mimicked by a Soluble Trimeric Immunogen

Summary Many broadly neutralizing antibodies (bnAbs) against HIV-1 recognize and/or penetrate the glycan shield on native, virion-associated envelope glycoprotein (Env) spikes. The same bnAbs also bind to recombinant, soluble trimeric immunogens based on the SOSIP design. While SOSIP trimers are close structural and antigenic mimics of virion Env, the extent to which their glycan structures resemble ones on infectious viruses is undefined. Here, we compare the overall glycosylation of gp120 and gp41 subunits from BG505 (clade A) virions produced in a lymphoid cell line with those from recombinant BG505 SOSIP trimers, including CHO-derived clinical grade material. We also performed detailed site-specific analyses of gp120. Glycans relevant to key bnAb epitopes are generally similar on the recombinant SOSIP and virion-derived Env proteins, although the latter do contain hotspots of elevated glycan processing. Knowledge of native versus recombinant Env glycosylation will guide vaccine design and manufacturing programs.

[1]  E. Go,et al.  Characterization of host-cell line specific glycosylation profiles of early transmitted/founder HIV-1 gp120 envelope proteins. , 2013, Journal of proteome research.

[2]  R. Desrosiers,et al.  Mapping the complete glycoproteome of virion-derived HIV-1 gp120 provides insights into broadly neutralizing antibody binding , 2016, Scientific Reports.

[3]  M. Crispin,et al.  Structural principles controlling HIV envelope glycosylation. , 2017, Current opinion in structural biology.

[4]  Dennis R Burton,et al.  Envelope glycans of immunodeficiency virions are almost entirely oligomannose antigens , 2010, Proceedings of the National Academy of Sciences.

[5]  D. Burton,et al.  A Broadly Neutralizing Antibody Targets the Dynamic HIV Envelope Trimer Apex via a Long, Rigidified, and Anionic β-Hairpin Structure , 2017, Immunity.

[6]  D. Burton,et al.  Unprecedented Role of Hybrid N-Glycans as Ligands for HIV-1 Broadly Neutralizing Antibodies. , 2018, Journal of the American Chemical Society.

[7]  J. Hoxie,et al.  Derivation and Characterization of a Simian Immunodeficiency Virus SIVmac239 Variant with Tropism for CXCR4 , 2009, Journal of Virology.

[8]  John R Yates,et al.  Global site-specific N-glycosylation analysis of HIV envelope glycoprotein , 2017, Nature Communications.

[9]  J. Hoxie,et al.  Envelope Glycoprotein Incorporation, Not Shedding of Surface Envelope Glycoprotein (gp120/SU), Is the Primary Determinant of SU Content of Purified Human Immunodeficiency Virus Type 1 and Simian Immunodeficiency Virus , 2002, Journal of Virology.

[10]  E. Go,et al.  Influences on the Design and Purification of Soluble, Recombinant Native-Like HIV-1 Envelope Glycoprotein Trimers , 2015, Journal of Virology.

[11]  John P. Moore,et al.  Molecular Architecture of the Cleavage-Dependent Mannose Patch on a Soluble HIV-1 Envelope Glycoprotein Trimer , 2016, Journal of Virology.

[12]  M. Crispin,et al.  Cell- and Protein-Directed Glycosylation of Native Cleaved HIV-1 Envelope , 2015, Journal of Virology.

[13]  L. Stamatatos,et al.  Germline‐targeting immunogens , 2017, Immunological reviews.

[14]  Yan Liu,et al.  Supersite of immune vulnerability on the glycosylated face of HIV-1 envelope glycoprotein gp120 , 2013, Nature Structural &Molecular Biology.

[15]  Michael S. Seaman,et al.  Complex-type N-glycan recognition by potent broadly neutralizing HIV antibodies , 2012, Proceedings of the National Academy of Sciences.

[16]  R. Warnke,et al.  Clinical and biologic characterization of T-cell neoplasias with rearrangements of chromosome 7 band q34. , 1988, Blood.

[17]  L. Stamatatos,et al.  Design and crystal structure of a native-like HIV-1 envelope trimer that engages multiple broadly neutralizing antibody precursors in vivo , 2017, The Journal of experimental medicine.

[18]  John P. Moore,et al.  A Next-Generation Cleaved, Soluble HIV-1 Env Trimer, BG505 SOSIP.664 gp140, Expresses Multiple Epitopes for Broadly Neutralizing but Not Non-Neutralizing Antibodies , 2013, PLoS pathogens.

[19]  D. Harvey.  Fragmentation of negative ions from carbohydrates: Part 3. Fragmentation of hybrid and complex N-linked glycans , 2005, Journal of the American Society for Mass Spectrometry.

[20]  R. Sanders,et al.  In vivo protection by broadly neutralizing HIV antibodies. , 2014, Trends in microbiology.

[21]  D. Burton,et al.  Incomplete Neutralization and Deviation from Sigmoidal Neutralization Curves for HIV Broadly Neutralizing Monoclonal Antibodies , 2015, PLoS pathogens.

[22]  D. Burton,et al.  The Glycan Shield of HIV Is Predominantly Oligomannose Independently of Production System or Viral Clade , 2011, PloS one.

[23]  James C Paulson,et al.  Structural delineation of a quaternary, cleavage-dependent epitope at the gp41-gp120 interface on intact HIV-1 Env trimers. , 2014, Immunity.

[24]  Ian A Wilson,et al.  Structural Constraints Determine the Glycosylation of HIV-1 Envelope Trimers. , 2015, Cell reports.

[25]  E. Go,et al.  Glycosylation Benchmark Profile for HIV-1 Envelope Glycoprotein Production Based on Eleven Env Trimers , 2017, Journal of Virology.

[26]  Martin A. Nowak,et al.  Antibody neutralization and escape by HIV-1 , 2003, Nature.

[27]  H. Widmer,et al.  Ion exchange and purification of carbohydrates on a Nafion® membrane as a new sample pretreatment for matrix‐assisted laser desorption/ionization mass spectrometry , 1995 .

[28]  M. Nussenzweig,et al.  Natively glycosylated HIV-1 Env structure reveals new mode for antibody recognition of the CD4-binding site , 2016, Nature Structural &Molecular Biology.

[29]  Lynn Morris,et al.  Evolution of an HIV glycan–dependent broadly neutralizing antibody epitope through immune escape , 2012, Nature Medicine.

[30]  Tongqing Zhou,et al.  Structural Basis for Broad and Potent Neutralization of HIV-1 by Antibody VRC01 , 2010, Science.

[31]  D. Burton,et al.  Glycans Function as Anchors for Antibodies and Help Drive HIV Broadly Neutralizing Antibody Development. , 2017, Immunity.

[32]  Weston B Struwe,et al.  Composition and Antigenic Effects of Individual Glycan Sites of a Trimeric HIV-1 Envelope Glycoprotein , 2016, Cell reports.

[33]  Daniel W. Kulp,et al.  Vaccine-Elicited Tier 2 HIV-1 Neutralizing Antibodies Bind to Quaternary Epitopes Involving Glycan-Deficient Patches Proximal to the CD4 Binding Site , 2015, PLoS pathogens.

[34]  D. Harvey.  Fragmentation of negative ions from carbohydrates: Part 2. Fragmentation of high-mannose N-linked glycans , 2005, Journal of the American Society for Mass Spectrometry.

[35]  J. Binley,et al.  A Recombinant Human Immunodeficiency Virus Type 1 Envelope Glycoprotein Complex Stabilized by an Intermolecular Disulfide Bond between the gp120 and gp41 Subunits Is an Antigenic Mimic of the Trimeric Virion-Associated Structure , 2000, Journal of Virology.

[36]  John P. Moore,et al.  Native‐like Env trimers as a platform for HIV‐1 vaccine design , 2017, Immunological reviews.

[37]  Yan Liu,et al.  A Potent and Broad Neutralizing Antibody Recognizes and Penetrates the HIV Glycan Shield , 2011, Science.

[38]  William R. Schief,et al.  Glycan clustering stabilizes the mannose patch of HIV-1 and preserves vulnerability to broadly neutralizing antibodies , 2015, Nature Communications.

[39]  J. Overbaugh,et al.  Human Immunodeficiency Virus Type 1 V1-V2 Envelope Loop Sequences Expand and Add Glycosylation Sites over the Course of Infection, and These Modifications Affect Antibody Neutralization Sensitivity , 2006, Journal of Virology.

[40]  Dennis R Burton,et al.  Broadly Neutralizing Antibodies to HIV and Their Role in Vaccine Design. , 2016, Annual review of immunology.

[41]  N. Haigwood,et al.  Use of broadly neutralizing antibodies for HIV‐1 prevention , 2017, Immunological reviews.

[42]  J. Overbaugh,et al.  Early development of broad neutralizing antibodies in HIV-1 infected infants , 2014, Nature Medicine.

[43]  E. Go,et al.  cGMP production and analysis of BG505 SOSIP.664, an extensively glycosylated, trimeric HIV‐1 envelope glycoprotein vaccine candidate , 2017, Biotechnology and bioengineering.

[44]  G. Debnath,et al.  D-101 HIV-1 neutralizing antibodies induced by native-like envelope trimers , 2016 .

[45]  B. Domon,et al.  A systematic nomenclature for carbohydrate fragmentations in FAB-MS/MS spectra of glycoconjugates , 1988, Glycoconjugate Journal.

[46]  Ian A Wilson,et al.  Structure and Immune Recognition of the HIV Glycan Shield. , 2018, Annual review of biophysics.

[47]  J. Mascola,et al.  HIV-1 envelope pseudotyped viral vectors and infectious molecular clones expressing the same envelope glycoprotein have a similar neutralization phenotype, but culture in peripheral blood mononuclear cells is associated with decreased neutralization sensitivity. , 2005, Virology.

[48]  Baoshan Zhang,et al.  Structural basis for diverse N-glycan recognition by HIV-1–neutralizing V1–V2–directed antibody PG16 , 2013, Nature Structural &Molecular Biology.