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Preparation, characterization and immunogenicity of HIV-1 related high-mannose oligosaccharides-CRM197 glycoconjugates

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Abstract

The dense glycan shield on the surface of human immunodeficiency virus type 1 (HIV-1) gp120 masks conserved protein epitopes and facilitates virus entry via interaction to glycan binding proteins on susceptible host cells. The broadly neutralizing monoclonal antibody 2G12 binds a cluster of high-mannose oligosaccharides on the gp120 subunit of HIV-1 Env protein. This oligomannose epitope is currently being considered for the design of a synthetic vaccine. The cluster nature of the 2G12 epitope suggests that a multivalent antigen presentation is important to develop a carbohydrate-based vaccine candidate. In this work we describe the development of neoglycoconjugates displaying clustered HIV-1 related oligomannose carbohydrates. We exploited flexible polyamidoamine (PAMAM) scaffold to generate four- and eight-valent sugar clusters of HIV-1-related oligomannose antigens Man4, Man6 and Man9. The multivalent presentation of oligomannoses increased the avidity of Man4 and Man9 to 2G12. The synthetic glycodendrons were then covalently coupled to the protein carrier CRM197, formulated with the adjuvant MF59, and used to immunize two animal species. Oligomannose-specific IgG antibodies were generated; however, the antisera failed to recognize recombinant HIV-1 gp120 proteins. We conclude that further structural vaccinology work is needed to identify an antigen presentation that closely matches in vivo the structure of the epitope mapped by 2G12.

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Abbreviations

CHO:

Carbohydrate

Man:

Mannose

RT:

Room temperature

vol:

Volume

GNL:

Galantus nivalis lectin

i.m.:

Intramuscular

sc.:

Subcutaneous

HSA:

Human serum albumin

IC50 :

Inhibitor concentration causing fifty % signal reduction

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Acknowledgements

We want to thank Novartis Animal Care facility for conduction of animal studies. We greatly appreciate the gift of PAMAM material from Prof. Giorgio Catelani of Pisa University (Italy).

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Authors and Affiliations

  1. Novartis Vaccines and Diagnostics, Research Center, Via Fiorentina 1, 53100, Siena, Italy

    Anna Kabanova, Roberto Adamo, Daniela Proietti, Francesco Berti, Marta Tontini, Rino Rappuoli & Paolo Costantino

  2. Department of Experimental Evolutionary Biology, University of Bologna, via Zamboni 33, 40126, Bologna, Italy

    Anna Kabanova

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  1. Anna Kabanova
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  2. Roberto Adamo
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  3. Daniela Proietti
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Correspondence to Paolo Costantino.

Online supplementary resources

Below is the link to the electronic supplementary material.

Online resource 1

ESI Q-TOF MS spectra of Man4-PAMAM4 and Man9-PAMAM8. (PDF 199 kb)

Online resource 2

1H-NMR 400 MHz spectra of the glycodendrons. (PDF 197 kb)

Online resource 3

Sensorgrams of the surface plasmon resonance inhibition assay. (PDF 374 kb)

Online resource 4

Synthetic high-mannose oligosaccharides characterization data including 1H-NMR 400 MHz profiles and MALDI-MS spectra. (PDF 317 kb)

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Kabanova, A., Adamo, R., Proietti, D. et al. Preparation, characterization and immunogenicity of HIV-1 related high-mannose oligosaccharides-CRM197 glycoconjugates. Glycoconj J 27, 501–513 (2010). https://doi.org/10.1007/s10719-010-9295-0

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