Humoral immune response induced with dengue virus-like particles serotypes 1 and 4 produced in silkworm

Abstract

Dengue is an arboviral disease, which threatens almost half the global population, and has emerged as the most significant of current global public health challenges. In this study, we prepared dengue virus-like particles (DENV-LPs) consisting of Capsid-premembrane-envelope (CprME) and premembrane-envelope (prME) polypeptides from serotype 1 and 4, which were expressed in the silkworms using Bombyx mori nucleopolyhedrovirus (BmNPV) bacmid. 1CprME, 1prME, 4CprME, and 4prME expressed proteins in hemolymph, and the molecular weight of the purified proteins was 55 kDa, respectively. The purified polypeptides formed spherical Dengue virus-like particles (DENV-LPs) with ~ 30–55 nm in diameter. The immunoelectron microscopy (IEM) images revealed antigens to the surface of a lipid bilayer of DENV-LPs. The heparin-binding assay shows a positive relationship between absorbance and E protein domain III (EDIII) quantity, which is supported by the isothermal titration calorimetry assay. This indicates a moderate binding affinity between heparin and DENV-LP. The high correlation between patient sera and DENV-LP reactivities revealed that these DENV-LPs shared similar epitopes with the natural dengue virus. IgG elicitation studies in mice have demonstrated that DENV-LPs/CPrMEs elicit a stronger immune response than DENV-LP/prMEs, which lends credence to this claim.

Display of multiple proteins on engineered canine parvovirus-like particles expressed in cultured silkworm cells and silkworm larvae

“…The purified and concentrated proteins from VP variants were subjected to TEM to confirm the VLP formation and morphology following our previous publications ( Suhaimi et al, 2019 ; Utomo et al, 2022 ). Briefly, a 20 μL protein solution drop was loaded onto the surface of a carbon film-supported copper grid (200 mesh, Nisshin Em Co. Ltd., Tokyo, Japan) and incubated at room temperature for 30 s. The grid was three-time washed with PBS and negatively stained with phosphotungstic acid (2% v/v).…”

An Overview of Recent Developments in the Application of Antigen Displaying Vaccine Platforms: Hints for Future SARS-CoV-2 VLP Vaccines

“…Dealing with viruses that have multiple variants or serotypes presents several challenges for vaccine development, such as the dengue viruses, for which a commercialized vaccine is still unavailable. Several studies have explored a potential VLP-based vaccine for developing the dengue vaccine, which may comprise several serotypes [ 43 , 44 ]. A recent study from Utomo et al [ 43 ] developed a dengue virus-like particle comprising a capsid-pre-membrane-envelope polypeptide from serotype 1–4 fused chemically, with which promising results in eliciting a robust immune response were seen in mice after vaccination.…”