Alphavirus capsid proteins self-assemble into core-like particles in insect cells: A promising platform for nanoparticle vaccine development.
The mosquito-borne chikungunya virus (CHIKV) causes arthritic diseases in humans, whereas the aquatic salmonid alphavirus (SAV) is associated with high mortality in aquaculture of salmon and trout. Using modern biotechnological approaches, promising vaccine candidates based upon highly immunogenic, enveloped virus-like particles (eVLPs) have been developed. However, the eVLP structure (core, lipid membrane, surface glycoproteins) is more complex than that of non-enveloped, protein-only VLPs, which are structurally and morphologically 'simple'. In order to develop an alternative to alphavirus eVLPs, in this paper we engineered recombinant baculovirus vectors to produce high levels of alphavirus core-like particles (CLPs) in insect cells by expression of the CHIKV and SAV capsid proteins. The CLPs localize in dense nuclear bodies within the infected cell nucleus and are purified through a rapid and scalable protocol involving cell lysis, sonication and low-speed centrifugation steps. Furthermore, an immunogenic epitope from the alphavirus E2 glycoprotein can be successfully fused to the N-terminus of the capsid protein without disrupting the CLP self-assembling properties. We propose that immunogenic epitope-tagged alphavirus CLPs produced in insect cells present a simple and perhaps more stable alternative to alphavirus eVLPs.
Laboratory of Virology, Wageningen University, Wageningen, The Netherlands.
Mia C
MC
Hikke
Laboratory of Virology, Wageningen University, Wageningen, The Netherlands.
Corinne
C
Geertsema
Laboratory of Virology, Wageningen University, Wageningen, The Netherlands.
Vincen
V
Wu
Laboratory of Virology, Wageningen University, Wageningen, The Netherlands.
Stefan W
SW
Metz
Laboratory of Virology, Wageningen University, Wageningen, The Netherlands.
Jan W
JW
van Lent
Laboratory of Virology, Wageningen University, Wageningen, The Netherlands.
Just M
JM
Vlak
Laboratory of Virology, Wageningen University, Wageningen, The Netherlands. gorben.pijlman@wur.nl.
Gorben P
GP
Pijlman
D000524:Alphavirus / Q000235:genetics* / Q000276:immunology; D000818:Animals; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism; D036022:Capsid Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D002467:Cell Nucleus; D015195:Drug Design; D000939:Epitopes / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000276:immunology; D061987:Sf9 Cells; D058425:Vaccines, Virus-Like Particle / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology*; D019065:Virus Assembly
D036022:Capsid Proteins; D000939:Epitopes; D011993:Recombinant Fusion Proteins; D058425:Vaccines, Virus-Like Particle; D014759:Viral Envelope Proteins
Alphavirus; Baculovirus expression; Core-like particle; Protein carrier; Subunit vaccine