Zika virus baculovirus-expressed envelope protein elicited humoral and cellular immunity in immunocompetent mice

AbstractZika virus (ZIKV) is a mosquito-borne virus that has a high risk of inducing Guillain–Barré syndrome and microcephaly in newborns. Because vaccination is considered the most effective strategy against ZIKV infection, we designed a recombinant vaccine utilizing the baculovirus expression system with two strains of ZIKV envelope protein (MR766, Env_M; ZBRX6, Env_Z). Animals inoculated with Env_M and Env_Z produced ZIKV-specific antibodies and secreted effector cytokines such as interferon-γ, tumor necrosis factor-α, and interleukin-12. Moreover, the progeny of immunized females had detectable maternal antibodies that protected them against two ZIKV strains (MR766 and PRVABC59) and a Dengue virus strain. We propose that the baculovirus expression system ZIKV envelope protein recombinant provides a safe and effective vaccine strategy.

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DETECTION OF ANTI-RETICULOENDOTHELIOSIS ANTIBODY BY ENZYME-LINKED IMMUNOSORBENT ASSAY USING ENVELOPE PROTEIN EXPRESSED IN BACULOVIRUS

Taiwan Veterinary Journal ◽

10.1142/s1682648515500249 ◽

2016 ◽

Vol 42 (03) ◽

pp. 165-170

Author(s):

Hui-Wen Chen ◽

Ting-Hsiang Hsiao ◽

Ching-Ho Wang

Keyword(s):

Immunosorbent Assay ◽

Envelope Protein ◽

Insect Cells ◽

Expression System ◽

Baculovirus Expression System ◽

Relative Sensitivity ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Baculovirus Expression ◽

Relative Specificity

Reticuloendotheliosis virus (REV) infects various animals including chickens, ducks, geese, pheasants, peafowl and other birds. REV causes tumor, runting disease syndrome and immunosuppression in infected birds. The purpose of this study was to develop a rapid, reliable and convenient method to detect anti-REV antibody. The REV envelope protein was expressed by baculovirus expression system and evaluated for the potential as an antigen in an enzyme linked-immunosorbent assay (ELISA). The env gene from a REV strain goose/3410/06 was cloned into bacmid vector and expressed in Sf9 insect cells. The expected recombinant envelope protein expressed in infected insect cells was demonstrated in western blot with size of 62 kDa, and purified for using as antigen in an ELISA. A total of 182 chicken serum samples were used to evaluate the suitability of the ELISA based on neutralization test as the gold standard. The results showed that the relative sensitivity and relative specificity of this ELISA were 88.5% and 97.7%, respectively. The present ELISA using baculovirus expression envelope protein can be used to detect anti-REV antibody in the field.

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Preparation of Chicken Anemia Virus (CAV) Virus-Like Particles and Chicken Interleukin-12 for Vaccine Development Using a Baculovirus Expression System

Pathogens ◽

10.3390/pathogens8040262 ◽

2019 ◽

Vol 8 (4) ◽

pp. 262

Author(s):

Ta-Yuan Tseng ◽

Yee-Chen Liu ◽

Yu-Chen Hsu ◽

Poa-Chun Chang ◽

Ming-Kun Hsieh ◽

...

Keyword(s):

Vaccine Development ◽

Expression System ◽

Baculovirus Expression System ◽

Chicken Anemia Virus ◽

Economic Losses ◽

Interleukin 12 ◽

Cell Mediated Immunity ◽

Single Chain ◽

Virus Like Particles ◽

Baculovirus Expression

Chicken infectious anemia (CIA) is a poultry disease that causes huge economic losses in the poultry industry worldwide. Commercially available CIA vaccines are derived from wild-type chicken anemia viruses (CAVs) by serial passage in cells or chicken embryos. However, these vaccinal viruses are not completely attenuated; therefore, they can be transmitted vertically and horizontally, and may induce clinical symptoms in young birds. In this study, we sought to eliminate these issues by developing a subunit vaccine exploiting the CAV structural proteins, engineering recombinant baculovirus-infected Spodoptera frugiperda (Sf9) cells that contained both the viral protein 1 (VP1) and VP2 of CAV. Moreover, we produced single-chain chicken interleukin-12 (chIL-12) in the same system, to serve as an adjuvant. The recombinant VP1 was recognized by chicken anti-CAV polyclonal antibodies in Western blotting and immunofluorescence assays, and the bioactivity of the recombinant chIL-12 was confirmed by stimulating interferon-γ (IFN-γ) secretion in chicken splenocytes. Furthermore, the ability of the recombinant VP1 to generate self-assembling virus-like particles (VLPs) was confirmed by transmission electron microscopy. Specific pathogen-free (SPF) chickens inoculated with VLPs and co-administered the recombinant chIL-12 induced high CAV-specific antibodies and cell-mediated immunity. Taken together, the VLPs produced by the baculovirus expression system have the potential to be a safe and effective CIA vaccine. Finally, we demonstrated the utility of recombinant chIL-12 as an adjuvant for poultry vaccine development.

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Expression of functional G protein beta gamma dimers of defined subunit composition using a baculovirus expression system.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)42178-3 ◽

1992 ◽

Vol 267 (19) ◽

pp. 13123-13126 ◽

Cited By ~ 4

Author(s):

S.G. Graber ◽

R.A. Figler ◽

V.K. Kalman-Maltese ◽

J.D. Robishaw ◽

J.C. Garrison

Keyword(s):

G Protein ◽

Expression System ◽

Baculovirus Expression System ◽

Subunit Composition ◽

Baculovirus Expression

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Production, processing and partial purification of functional G protein βγ subunits in baculovirus-infected insect cells

Biochemical Journal ◽

10.1042/bj2860677 ◽

1992 ◽

Vol 286 (3) ◽

pp. 677-680 ◽

Cited By ~ 21

Author(s):

J D Robishaw ◽

V K Kalman ◽

K L Proulx

Keyword(s):

G Protein ◽

G Proteins ◽

Insect Cells ◽

Expression System ◽

Baculovirus Expression System ◽

Partial Purification ◽

Baculovirus Expression ◽

Gamma Subunits ◽

Infected Insect ◽

Beta 2

As a result of the inability to resolve the heterogeneous mixture of G protein beta gamma subunits present in tissues, it has not been possible to compare different beta gamma subunits of the G proteins in terms of their proposed roles in receptor-effector coupling. This study was undertaken to establish the utility of the baculovirus expression system in producing homogeneous beta gamma subunits of defined composition for the comparative analysis of these subunits in reconstitution systems. In this study we report the expression, and appropriate post-translational processing, of recombinant beta 2, gamma 2 and gamma 3 subunits. In addition, we show that the recombinant beta gamma subunits can be readily purified, and can functionally interact with the alpha subunits of the G proteins.

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