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

2022 ◽  
Vol 12 (1) ◽  
Author(s):  
Minna Shin ◽  
Kiju Kim ◽  
Hyo-Ji Lee ◽  
Rangyeon Lee ◽  
Yu-Jin Jung ◽  
...  

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.

2016 ◽  
Vol 42 (03) ◽  
pp. 165-170
Author(s):  
Hui-Wen Chen ◽  
Ting-Hsiang Hsiao ◽  
Ching-Ho Wang

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.


Pathogens ◽  
2019 ◽  
Vol 8 (4) ◽  
pp. 262
Author(s):  
Ta-Yuan Tseng ◽  
Yee-Chen Liu ◽  
Yu-Chen Hsu ◽  
Poa-Chun Chang ◽  
Ming-Kun Hsieh ◽  
...  

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.


1992 ◽  
Vol 267 (19) ◽  
pp. 13123-13126 ◽  
Author(s):  
S.G. Graber ◽  
R.A. Figler ◽  
V.K. Kalman-Maltese ◽  
J.D. Robishaw ◽  
J.C. Garrison

1992 ◽  
Vol 286 (3) ◽  
pp. 677-680 ◽  
Author(s):  
J D Robishaw ◽  
V K Kalman ◽  
K L Proulx

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.


2016 ◽  
Vol 11 ◽  
pp. 86-89 ◽  
Author(s):  
Tsuyoshi Ikehara ◽  
Shihoko Nakashima ◽  
Junichi Nakashima ◽  
Tsubasa Kinoshita ◽  
Takeshi Yasumoto

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