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Characterization and Immunogenicity of HIV Envelope gp140 Zera^® Tagged Antigens.

ABSTRACT: HIV-1 envelope glycoprotein (Env) remains the most relevant target for the elicitation of functional antibodies to HIV by vaccination. However, soluble Env antigens often do not elicit the desired immune responses. Delivering subunit antigens on particulate nanoparticles is an established approach to improve their immunogenicity. In this study the sequence encoding Zera^®, a proline-rich domain derived from the ?-zein storage protein, was fused to either the C- or N-terminus of the superinfecting HIV-1 CAP256 gp140 envelope: Zera^® generally induces the formation of protein bodies (PBs), which can significantly improve both the immunogenicity and yields of the partner protein. The expression of gp140-Zera^® and Zera^®-gp140 (N- and C-terminal fusions respectively) in mammalian cells was confirmed by western blot analysis and immunostaining. However, isopycnic ultracentrifugation showed that neither gp140-Zera^® nor Zera^®-gp140 accumulated in characteristic electron-dense PBs. gp140-Zera^® elicited higher binding antibody titers in rabbits to autologous gp140 and V1V2 scaffold than Zera^®-gp140. Rabbit anti-gp140-Zera^® sera also had significantly higher Tier 1A neutralizing antibody titers than anti-Zera^®-gp140 sera. Neither gp140-Zera^® nor Zera^®-gp140-specific sera neutralized Tier 1B or autologous Tier 2 viruses. These results showed that HIV-1 gp140 tagged with Zera^® at either the N- or C-termini elicited high titers of gp140 and V1V2 binding antibodies, and low levels of Tier 1 neutralizing antibodies in rabbits.

SUBMITTER: Ximba P

PROVIDER: S-EPMC7160593 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Characterization and Immunogenicity of HIV Envelope gp140 Zera<sup>®</sup> Tagged Antigens.

Ximba Phindile P Chapman Rosamund R Meyers Ann E AE Margolin Emmanuel E van Diepen Michiel T MT Williamson Anna-Lise AL Rybicki Edward P EP

Frontiers in bioengineering and biotechnology 20200409

HIV-1 envelope glycoprotein (Env) remains the most relevant target for the elicitation of functional antibodies to HIV by vaccination. However, soluble Env antigens often do not elicit the desired immune responses. Delivering subunit antigens on particulate nanoparticles is an established approach to improve their immunogenicity. In this study the sequence encoding Zera<sup>®</sup>, a proline-rich domain derived from the γ-zein storage protein, was fused to either the C- or N-terminus of the sup ...[more]

PMID: 32328488

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Project description:UnlabelledThe human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein (Env) trimer, which consists of the gp120 and gp41 subunits, is the focus of multiple strategies for vaccine development. Extensive Env glycosylation provides HIV-1 with protection from the immune system, yet the glycans are also essential components of binding epitopes for numerous broadly neutralizing antibodies. Recent studies have shown that when Env is isolated from virions, its glycosylation profile differs significantly from that of soluble forms of Env (gp120 or gp140) predominantly used in vaccine discovery research. Here we show that exogenous membrane-anchored Envs, which can be produced in large quantities in mammalian cells, also display a virion-like glycan profile, where the glycoprotein is extensively decorated with high-mannose glycans. Additionally, because we characterized the glycosylation with a high-fidelity profiling method, glycopeptide analysis, an unprecedented level of molecular detail regarding membrane Env glycosylation and its heterogeneity is presented. Each glycosylation site was characterized individually, with about 500 glycoforms characterized per Env protein. While many of the sites contain exclusively high-mannose glycans, others retain complex glycans, resulting in a glycan profile that cannot currently be mimicked on soluble gp120 or gp140 preparations. These site-level studies are important for understanding antibody-glycan interactions on native Env trimers. Additionally, we report a newly observed O-linked glycosylation site, T606, and we show that the full O-linked glycosylation profile of membrane-associated Env is similar to that of soluble gp140. These findings provide new insight into Env glycosylation and clarify key molecular-level differences between membrane-anchored Env and soluble gp140.ImportanceA vaccine that protects against human immunodeficiency virus type 1 (HIV-1) infection should elicit antibodies that bind to the surface envelope glycoproteins on the membrane of the virus. The envelope glycoproteins have an extensive coat of carbohydrates (glycans), some of which are recognized by virus-neutralizing antibodies and some of which protect the virus from neutralizing antibodies. We found that the HIV-1 membrane envelope glycoproteins have a unique pattern of carbohydrates, with many high-mannose glycans and also, in some places, complex glycans. This pattern was very different from the carbohydrate profile seen for a more easily produced soluble version of the envelope glycoprotein. Our results provide a detailed characterization of the glycans on the natural membrane envelope glycoproteins of HIV-1, a carbohydrate profile that would be desirable to mimic with a vaccine.

Dataset Information

Characterization and Immunogenicity of HIV Envelope gp140 Zera^® Tagged Antigens.

Publications

Characterization and Immunogenicity of HIV Envelope gp140 Zera<sup>®</sup> Tagged Antigens.

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Dataset Information

Characterization and Immunogenicity of HIV Envelope gp140 Zera® Tagged Antigens.

Publications

Characterization and Immunogenicity of HIV Envelope gp140 Zera<sup>®</sup> Tagged Antigens.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Characterization and Immunogenicity of HIV Envelope gp140 Zera^® Tagged Antigens.