Abstract
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.
Highlights
Flaviviruses like dengue virus (DENV) and West Nile virus (WNV) have established strongholds in many parts of tropical and sub-tropical countries world-wide
The fact that virus-like particles (VLPs) were purified by two ultracentrifugation steps, velocity ultracentrifugation through a sucrose cushion, followed by isopycnic ultracentrifugation in a sucrose-D2O preformed gradient, and that domain III (DIII)-receptor of IgE (RIgE) remained associated with Gag polyprotein throughout the purification procedure suggested that retroviral VLPs were pseudotyped with the human glycoprotein CD16-RIgE
The choice of the strategy of VLP-display of the structural DIII domain of flavivirus envelope glycoprotein for potential vaccine antigen was based on the following considerations
Summary
Flaviviruses like dengue virus (DENV) and West Nile virus (WNV) have established strongholds in many parts of tropical and sub-tropical countries world-wide. They are among the most important agents of re-emerging diseases. VLPs consisting only of viral envelope glycoproteins have been isolated, such as the subviral particles formed by hepatitis B virus (HBV) envelope glycoprotein S, or by chimeric HBV and hepatitis C virus (HCV) envelope proteins [6,7]. In the case of the influenza virus, VLPs produced by recombinant baculovirus-infected insect cells consisted of two membrane-associated proteins, hemagglutinin and matrix protein [8]
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