Development and immunological assessment of VLP-based immunogens exposing the membrane-proximal region of the HIV-1 gp41 protein

Journal of Biomedical Science

Ralf Wagner²  Jonathan L Heeney³  Ruth Kapzan⁴  Alexander Kliche²  Paul Tonks³  Thomas D Benen¹ 

[1] Current address: Malvern Instruments GmbH, Herrenberg, Germany;Molecular Microbiology and Gene Therapy Unit, Institute of Medical Microbiology and Hygiene, University of Regensburg, Franz-Josef-Strauss-Allee 11, Regensburg, 93053, Germany;Department of Veterinary Medicine, University of Cambridge, Cambridge, UK;Current address: GeneArt AG / LifeTechnologies Inc, Regensburg, Germany

关键词: Neutralizing antibodies;    Immunization study;    Vaccine;    VLPs;    gp41;    HIV-1;   

Others  :  1146516
DOI  :  10.1186/s12929-014-0079-x

 received in 2014-02-22, accepted in 2014-08-11,  发布年份 2014

【 摘 要 】

Background

The membrane-proximal external region (MPER) of HIV-1 gp41 is particularly conserved and target for the potent broadly neutralizing monoclonal antibodies (bnMAbs) 2F5, 4E10 and 10E8. Epitope focusing and stabilization present promising strategies to enhance the quality of immune responses to specific epitopes.

Results

The aim of this work was to design and evaluate novel immunogens based on the gp41 MPER with the potential to elicit cross-clade neutralizing antibodies. For that purpose, gp41 was truncated N-terminally in order to dispose immunodominant, non-neutralizing sites and enhance the exposure of conserved regions. To stabilize a trimeric conformation, heterologous GCN4 and HA2 zipper domains were fused based on an in silico “best-fit” model to the protein’s amino terminus. Cell surface exposure of resulting proteins and their selective binding to bnMAbs 2F5 and 4E10 could be shown by cytometric analyses. Incorporation into VLPs and preservation of antigenic structures were verified by electron microscopy, and the oligomeric state was successfully stabilized by zipper domains. These gp41 immunogens were evaluated for antigenicity in an immunization study in rabbits primed with homologous DNA expression plasmids and boosted with virus-like particle (VLP) proteins. Low titers of anti-MPER antibodies were measured by IgG ELISA, and low neutralizing activity could be detected against a clade C and B viral isolate in sera.

Conclusions

Thus, although neutralizing titers were very moderate, induction of cross-clade neutralizing antibodies seems possible following immunization with MPER-focusing immunogens. However, further refinement of MPER presentation and immunogenicity is clearly needed to induce substantial neutralization responses to these epitopes.

【 授权许可】

   
2014 Benen et al.; licensee BioMed Central

【 图 表 】

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