{"id":"doi:10.1371/journal.pone.0215031","kind":"article","title":"Delivery of self-amplifying RNA vaccines in in vitro reconstituted virus-like particles","authors":["Biddlecome A","Habte HH","McGrath KM","Sambanthamoorthy S","Wurm M","Sykora MM","Knobler CM","Lorenz IC","Lasaro M","Elbers K","Gelbart WM"],"author_count":11,"year":2019,"published":"2019-06-04","venue":"PloS one","cited_by":54,"doi":"10.1371/journal.pone.0215031","arxiv_id":null,"nct_id":null,"source_url":"https://doi.org/10.1371/journal.pone.0215031","archive_url":"https://space-kitty.com/papers/doi:10.1371/journal.pone.0215031","updated":null,"pmid":"31163034","pmcid":"PMC6548422","openalex_id":null,"pdf_url":"https://europepmc.org/articles/PMC6548422/pdf","abstract":"Many mRNA-based vaccines have been investigated for their specific potential to activate dendritic cells (DCs), the highly-specialized antigen-presenting cells of the immune system that play a key role in inducing effective CD4+ and CD8+ T-cell responses. In this paper we report a new vaccine/gene delivery platform that demonstrates the benefits of using a self-amplifying (\"replicon\") mRNA that is protected in a viral-protein capsid. Purified capsid protein from the plant virus Cowpea Chlorotic Mottle Virus (CCMV) is used to in vitro assemble monodisperse virus-like particles (VLPs) containing reporter proteins (e.g., Luciferase or eYFP) or the tandem-repeat model antigen SIINFEKL in RNA gene form, coupled to the RNA-dependent RNA polymerase from the Nodamura insect virus. Incubation of immature DCs with these VLPs results in increased activation of maturation markers - CD80, CD86 and MHC-II - and enhanced RNA replication levels, relative to incubation with unpackaged replicon mRNA. Higher RNA uptake/replication and enhanced DC activation were detected in a dose-dependent manner when the CCMV-VLPs were pre-incubated with anti-CCMV antibodies. In all experiments the expression of maturation markers correlates with the RNA levels of the DCs. Overall, these studies demonstrate that: VLP protection enhances mRNA uptake by DCs; coupling replicons to the gene of interest increases RNA and protein levels in the cell; and the presence of anti-VLP antibodies enhances mRNA levels and activation of DCs in vitro. Finally, preliminary in vivo experiments involving mouse vaccinations with SIINFEKL-replicon VLPs indicate a small but significant increase in antigen-specific T cells that are doubly positive for IFN and TFN induction.","abstract_license":"CC-BY (article license as recorded by Europe PMC; abstract reproduced unmodified apart from markup removal)","license":"CC-BY","is_open_access":true,"retracted":false,"peer_reviewed":null,"categories":[],"details":null,"provenance":"europepmc","attribution":"Abstract of \"Delivery of self-amplifying RNA vaccines in in vitro reconstituted virus-like particles\" by Biddlecome A, Habte HH, McGrath KM et al., https://doi.org/10.1371/journal.pone.0215031, used under its Creative Commons Attribution license (CC-BY); markup removed, otherwise unmodified.","topics":[{"slug":"mrna-cancer-vaccines","name":"mRNA Therapeutics & Cancer Vaccines","domain_slug":"medicine"}],"cited_in_dossiers":[]}