Open Access Highly Accessed Research

A novel platform for virus-like particle-display of flaviviral envelope domain III: induction of Dengue and West Nile virus neutralizing antibodies

Anthony JS Chua12, Cyrielle Vituret34, Melvin LC Tan1, Gaëlle Gonzalez34, Pierre Boulanger3, Mah-Lee Ng125* and Saw-See Hong346*

Author Affiliations

1 Flavivirology Laboratory, Department of Microbiology, National University of Singapore, 5 Science Drive 2, Singapore, 117597, Singapore

2 NUS Graduate School for Integrative Sciences and Engineering, National University of Singapore, 28 Medical Drive, Singapore, 117456, Singapore

3 University Lyon I & UMS-3444 Biosciences Gerland-Lyon Sud, 50, avenue Tony Garnier, Lyon, 69366, France

4 Retroviruses and Comparative Pathology, Université Lyon I & INRA UMR-754, 50, avenue Tony Garnier, Lyon Cedex 07, 69366, France

5 Flavivirology Laboratory, Department of Microbiology, National University Health System, 1E, Kent Ridge Road, Singapore, 119228, Singapore

6 Institut National de la Santé et de la Recherche Médicale, 101, rue de Tolbiac, Paris, 75013, France

For all author emails, please log on.

Virology Journal 2013, 10:129  doi:10.1186/1743-422X-10-129

Published: 24 April 2013


CD16-RIgE is a chimeric human membrane glycoprotein consisting of the CD16 ectodomain fused to the transmembrane domain and cytoplasmic tail of the gamma chain of the high affinity receptor of IgE (RIgE). Coexpression of CD16-RIgE and HIV-1 Pr55Gag polyprotein precursor (Pr55GagHIV) in insect cells resulted in the incorporation of CD16-RIgE glycoprotein into the envelope of extracellular virus-like particles (VLPs), a phenomenon known as pseudotyping. Taking advantage of this property, we replaced the CD16 ectodomain of CD16-RIgE by the envelope glycoprotein domain III (DIII) of dengue virus serotype 1 (DENV1) or West Nile virus Kunjin (WNVKun). The two resulting chimeric proteins, DIII-DENV1-RIgE and DIII-WNVKun-RIgE, were addressed to the plasma membrane, exposed at the surface of human and insect cells, and incorporated into extracellular VLPs when coexpressed with Pr55GagHIV in insect cells. The DIII domains were accessible at the surface of retroviral VLPs, as shown by their reactivity with specific antibodies, and notably antibodies from patient sera. The DIII-RIgE proteins were found to be incorporated in VLPs made of SIV, MLV, or chimeric MLV-HIV Gag precursors, indicating that DIII-RIgE could pseudotype a wide variety of retroviral VLPs. VLP-displayed DIII were capable of inducing specific neutralizing antibodies against DENV and WNV in mice. Although the neutralization response was modest, our data confirmed the capability of DIII to induce a flavivirus neutralization response, and suggested that our VLP-displayed CD16-RIgE-based platform could be developed as a vaccine vector against different flaviviruses and other viral pathogens.

Flavivirus envelope glycoprotein; Domain III; Retroviral Gag; Virus-like particles (VLPs); Pseudotyping; VLP-display; CD16/FcϵRIγ chimera; Recombinant baculovirus