Production of Japanese encephalitis virus-like particles in insect cells

Yamaji, Hideki; Konishi, Eiji

doi:10.4161/bioe.24514

Cited by 20 publications

(21 citation statements)

References 35 publications

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“…4(b), E protein expressed either in the presence or absence of prM peaked in the middle fractions, confirming that in the absence of prM, E protein was secreted in a particulate form. This is in addition to it being co-expressed in the form of prM-E, as reported previously (Yamaji & Konishi, 2013;Yamaji et al, 2012). It was noteworthy that the distribution of E antigen was not correlated with the distribution of GP64 and VP39, which represented the envelope and the capsid of the baculovirus, respectively, suggesting that the presence of the particulate form of JEV E was not simply due to incorporation into baculovirus particles.…”

Section: Jev E Forms Vlps Independently Of Prmsupporting

confidence: 84%

“…Since our data showed that little E protein was detected on the surface of vAc-prME infected cells, and since it has been also been established that the co-expression of JEV prM and E protein generates VLPs (Yamaji & Konishi, 2013;Yamaji et al, 2012), we therefore theorize that the observed syncytium in vAc-prME infected cells were induced by these VLPs in a 'fusion-from-without' manner (Bratt & Gallaher, 1969).…”

Section: Jev E Forms Vlps Independently Of Prmmentioning

confidence: 54%

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Glycoprotein E of the Japanese encephalitis virus forms virus-like particles and induces syncytia when expressed by a baculovirus

Yin

Wang

et al. 2015

Journal of General Virology

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The prM glycoprotein is thought to be a chaperone for the proper folding, membrane association and assembly of the envelope protein (E) of flaviviruses. The prM-E and E proteins of the Japanese encephalitis virus (JEV) were expressed in insect cells using both the baculovirus-expression system and the transient expression method. Protein expression was analysed by Western blotting and the cytopathic effect was observed by microscopy. In the baculovirus-expression system the E protein, with or without the prM protein, induced syncytial formation in Sf9 cells. Transient expression of prM-E also induced syncytia in Sf9 cells. Immunofluorescence revealed that in presence of prM, E proteins were endoplasmic reticulum-like in distribution, while in the absence of prM, E proteins were located on the cell surface. Sucrose gradient sedimentation and Western blot analysis indicated that the E protein expressed with or without the prM protein was secreted into the culture medium in particulate form. The formation of virus-like particles (VLPs) in the medium was confirmed by electron microscopy and immunoelectron microscopy. The results suggest that the E protein of JEV in the absence of prM, retained its fusion ability, by either cell surface expression or formation of VLPs. Moreover, based on the observation that co-expression of prM-E in Sf9 cells induced considerable syncytial formation, a novel, safe and simple antiviral screening approach is proposed for studying inhibitory antibodies, peptides or small molecules targeting the JEV E protein.

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Section: Jev E Forms Vlps Independently Of Prmsupporting

confidence: 84%

Section: Jev E Forms Vlps Independently Of Prmmentioning

confidence: 54%

Glycoprotein E of the Japanese encephalitis virus forms virus-like particles and induces syncytia when expressed by a baculovirus

Yin

Wang

et al. 2015

Journal of General Virology

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“…45,46 Some recombinant viral antigens can spontaneously assemble into virus-like particles (VLPs) which are multiprotein structures without the incorporation of a viral genome. 47,48 VLPs is a potentially safe subunit vaccine that can induce immune responses similar to those prompted by natural viral infection. 49 VLPs present repetitive high-density displays of epitopes and exhibit excellent adjuvant properties capable of inducing strong immune responses.…”

Section: Recombinant Protein Subunit Vaccinesmentioning

confidence: 99%

Recent advances in the production of recombinant subunit vaccines inPichia pastoris

2016

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Recombinant protein subunit vaccines are formulated using defined protein antigens that can be produced in heterologous expression systems. The methylotrophic yeast Pichia pastoris has become an important host system for the production of recombinant subunit vaccines. Although many basic elements of P. pastoris expression system are now well developed, there is still room for further optimization of protein production. Codon bias, gene dosage, endoplasmic reticulum protein folding and culture condition are important considerations for improved production of recombinant vaccine antigens. Here we comment on current advances in the application of P. pastoris for the synthesis of recombinant subunit vaccines.

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“…In addition, TBEV, JEV, WNV, and DENV SVPs were produced in CHO or HEK293T cells by expression of prM and E [26–29]. Other studies have shown that DENV, JEV, and WNV SVPs (based on either C‐prM‐E or prM‐E expression) could also be efficiently produced in yeast [30] or in insect cells, either transiently [31–33] or by using the baculovirus expression system [34, 35].…”

Section: Biotechnological Production Of Arboviral Evlps and Their Vmentioning

confidence: 99%

Enveloped virus‐like particles as vaccines against pathogenic arboviruses

Pijlman

2015

Biotechnology Journal

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Arthropod-borne arboviruses form a continuous threat to human and animal health, but few arboviral vaccines are currently available. Advances in expression technology for complex, enveloped virus-like particles (eVLPs) create new opportunities to develop potent vaccines against pathogenic arboviruses. In this short review, I highlight the successes and challenges in eVLP production for members of the three major arbovirus families: Flaviviridae (e.g., dengue, West Nile, Japanese encephalitis); Bunyaviridae (e.g., Rift Valley fever); and Togaviridae (e.g., chikungunya). The results from pre-clinical testing will be discussed as well as specific constraints to the large-scale manufacture and purification of eVLPs, which are complex assemblies of membranes and viral glycoproteins. Insect cells emerge as ideal substrates for correct arboviral glycoprotein folding and posttranslational modification to yield high quality eVLPs. Furthermore, baculovirus expression in insect cell culture is scalable and has a proven safety record in industrial human and veterinary vaccine manufacturing. In conclusion, eVLPs produced in insect cells using modern biotechnology have a realistic potential to be used in novel vaccines against arboviral diseases.

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Production of Japanese encephalitis virus-like particles in insect cells

Cited by 20 publications

References 35 publications

Glycoprotein E of the Japanese encephalitis virus forms virus-like particles and induces syncytia when expressed by a baculovirus

Glycoprotein E of the Japanese encephalitis virus forms virus-like particles and induces syncytia when expressed by a baculovirus

Recent advances in the production of recombinant subunit vaccines inPichia pastoris

Enveloped virus‐like particles as vaccines against pathogenic arboviruses

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