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

Doddy Irawan Setyo Utomo,Sabar Pambudi,Enoch Y Park

doi:10.1186/s13568-022-01353-6

Doddy Irawan Setyo Utomo, Sabar Pambudi + Show 1 more

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https://doi.org/10.1186/s13568-022-01353-6

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Journal: AMB Express	Publication Date: Jan 31, 2022
Citations: 2	License type: open-access

Affiliation: Shizuoka University

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.

Highlights

Dengue fever is a significant public health issue that has been reported in the Americas, Africa, Southeast Asia, Europe, the Western Pacific, and the Eastern Mediterranean
Expression of 1CprME, 1prME, 4CprME and 4prME polypeptides in silkworm Dengue virus (DENV) structural proteins are composed of a C, and two membrane proteins, prM and E translated at the beginning of the polyprotein in the order C-prM-E
Heparin binds to Dengue viruslike particle (DENV-LP) with Dissociation constant (KD) values of 197–952 nM and Gibbs free energy (ΔG) values from – 8.8 to – 9.3 kcal mol–1 (Table 2)

Summary

Introduction

Dengue fever is a significant public health issue that has been reported in the Americas, Africa, Southeast Asia, Europe, the Western Pacific, and the Eastern Mediterranean. This arboviral disease is endemic in more than 100 countries, and approximately 96 million infected individuals have symptoms of varying severities. Despite numerous attempts, identifying the best candidate for a dengue vaccine continues to be a difficult task due to some critical factors that must be considered (Bhatt et al 2021; Redoni et al 2020). DENV belongs to the Flaviviridae family and Flavivirus genus. It has four antigenically distinct serotypes and antigenically similar yet genetically diverse. DENV is an enveloped virus with a single positive-strand RNA genome that encodes three structural proteins, Capsid (C), premembrane (prM), and envelope (E), as well as seven nonstructural proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5) (Redoni et al 2020; Yousaf et al 2018)

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