Abstract
To develop monovalent dengue virus-like particle for serotype 3 (DENV-LP/3), we prepared and expressed two structural polyprotein constructs using silkworm and Bm5 cells: DENV-3 Capsid-premembrane-envelope (DENV-3CprME) and premembrane-envelope (DENV-3prME). The expressed PA-tagged 3CprME and 3prME polypeptides were partially purified by PA-tag affinity chromatography and had molecular weights of 85 and 75 kDa, respectively. Expressed proteins were separately verified using the following primary antibodies: the anti-PA tag antibody, DENV premembrane polyclonal antibody, and DENV envelope polyclonal antibody. Transmission electron microscopy revealed that these DENV-3CprME and 3prME formed rough, spherical DENV-LPs (DENV-LP/3CprME and DENV-LP/3prME), respectively, with a diameter of 30–55 nm. The heparin-binding assay demonstrated that these DENV-LPs contained the envelope protein domain III on their surfaces. Both DENV-LPs showed an affinity to sera from human dengue patients and immunized mice. Immunization of mice with DENV-LP/3prME significantly induced the level of antibodies compared with DENV-LP/3CprME. These results indicate that DENV-LP/3prME is suitable as a vaccine candidate compared with DENV-LP/3CprME.
Highlights
Dengue virus (DENV) is a single-stranded RNA arbovirus that has caused a global epidemic, which is most prominent in tropical and subtropical regions of the Americas, Asia, Africa, and the Pacific Islands
Expression of 3CprME and 3prME polypeptides in silkworm The DENV structural proteins consist of the C, prM, and E proteins, which are translated in the same order at the beginning of the polyprotein
The constructed Bombyx mori nucleopolyhedrovirus (BmNPV)/3CprME (Fig. 1a) and /3prME (Fig. 1b) bacmids were injected into silkworm larvae, and the silkworm hemolymph and fat body were collected at 5 dpi
Summary
Dengue virus (DENV) is a single-stranded RNA arbovirus that has caused a global epidemic, which is most prominent in tropical and subtropical regions of the Americas, Asia, Africa, and the Pacific Islands. The number of cases of dengue has been registered by the World Health Organization (WHO) since the 1950s, In 2016, the WHO estimated a global case rate of over 3.34 million cases a year, compared with a case rate of < 1000 cases a year in the 1950s. Primary infection by each of the four serotypes results in long-term serotype-specific immunity and a six-month period of partial immunity to other serotypes. After this partial immunity weakens, the person becomes vulnerable to infection by the other three DENV serotypes. This secondary infection may result in the antibody-dependent enhancement/clinical manifestations (Murrell et al 2011)
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