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BMC Biotechnology

, 12:80

Applied immunology

Abstract

BackgroundArenavirus matrix protein Z plays an important role in virus budding and is able to generate enveloped virus-like-particles VLPs in absence of any other viral proteins.
In these VLPs, Z protein is associated to the plasma membrane inner surface by its myristoyl residue.
Budding induction and vesicle formation properties can be exploited to generate enveloped VLPs platform.
These structures can be designed to carry specific antigen in the inner side or on the surface of VLPs.

Vaccines based on VLPs are a highly effective type of subunit vaccines that mimic the overall structure of virus particles in absence of viral nucleic acid, being noninfectious.

In this work we assayed the capacity of Junin Z protein to produce VLPs carrying the green fluorescent protein eGFP, as a model antigen.

ResultsIn this report the Junin Z protein ability to produce VLPs from 293T cells and its capacity to deliver a specific antigen eGFP fused to Z was evaluated.
Confocal microscopy showed a particular membrane bending in cells expressing Z and a spot welded distribution in the cytoplasm.
VLPs were detected by TEM transmission electron microscopy and were purified from cell supernatant.
The proteinase protection assay demonstrated the VLPs integrity and the absence of degradation of the fused antigen, thus indicating its internal localization.
Finally, immunization of mice with purified VLPs produced high titres of anti-eGFP antibodies compared to the controls.

ConclusionsIt was proved that VLPs can be generated from cells transfected with a fusion Junin virus Z-eGFP protein in absence of any other viral protein, and the capacity of Z protein to support fusions at the C-terminal, without impairing its budding activity, allowing vehiculization of specific antigens into VLPs.

KeywordsVirus-like particlesAntigen deliveryArenavirusZ proteinAbbreviationsCMVCytomegalovirus

MVBMultivesicular bodies

pDCPlasmacytoid dendritic cells

PMLPromyelocytic Leukemia Protein

TEMTransmission electron microscopy

VLPVirus-like-particles.

Electronic supplementary materialThe online version of this article doi:10.1186-1472-6750-12-80 contains supplementary material, which is available to authorized users.

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Autor: CristinaSBorio - MarcosFBilen - MarceloHArgelles - SandraEGoi - JavierAIserte - GracielaGlikmann - MarioELozano

Fuente: https://link.springer.com/



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