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Self-Replicating RNA Vaccine Delivery to Dendritic Cells

  • Thomas DémoulinsEmail author
  • Pavlos C. Englezou
  • Panagiota Milona
  • Nicolas Ruggli
  • Nicola Tirelli
  • Chantal Pichon
  • Cédric Sapet
  • Thomas Ebensen
  • Carlos A. Guzmán
  • Kenneth C. McCullough
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1499)

Abstract

Most current vaccines are either inactivated pathogen-derived or protein/peptide-based, although attenuated and vector vaccines have also been developed. The former induce at best moderate protection, even as multimeric antigen, due to limitations in antigen loads and therefore capacity for inducing robust immune defense. While attenuated and vector vaccines offer advantages through their replicative nature, drawbacks and risks remain with potential reversion to virulence and interference from preexisting immunity. New advances averting these problems are combining self-amplifying replicon RNA (RepRNA) technology with nanotechnology. RepRNA are large self-replicating RNA molecules (12–15 kb) derived from viral genomes defective in at least one structural protein gene. They provide sustained antigen production, effectively increasing vaccine antigen payloads over time, without the risk of producing infectious progeny. The major limitation with RepRNA is RNase-sensitivity and inefficient uptake by dendritic cells (DCs)—absolute requirements for efficacious vaccine design. We employed biodegradable delivery vehicles to protect the RepRNA and promote DC delivery. Encapsulating RepRNA into chitosan nanoparticles, as well as condensing RepRNA with polyethylenimine (PEI), cationic lipids, or chitosans, has proven effective for delivery to DCs and induction of immune responses in vivo.

Keywords

Replicon-RNA Self-replicating vaccine Universal influenza vaccine Dendritic cell delivery Chitosan nanoparticles Polyplexes Cationic lipids 

Notes

Acknowledgement

We thank Markus Gerber and Samira Locher for their help and input with the RepRNA technology, and Brigitte Herrmann for helping with the DC studies. We are also grateful to Patrick Midoux and Laure Magrangeas-Janot for help with the polyplex technology, Olivier Zelphati and Florent Poulhes for help with the lipoplex technology, and Kai Schulze for the adaptive immune response profiling. The work was funded by the Marie Curie IAPP Project Replixcel (251420) and the EU FP7 Project UniVax (HEALTH-F3-2013-60173).

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Thomas Démoulins
    • 1
    Email author
  • Pavlos C. Englezou
    • 1
  • Panagiota Milona
    • 1
  • Nicolas Ruggli
    • 1
  • Nicola Tirelli
    • 2
  • Chantal Pichon
    • 3
  • Cédric Sapet
    • 4
  • Thomas Ebensen
    • 5
  • Carlos A. Guzmán
    • 5
  • Kenneth C. McCullough
    • 1
  1. 1.Institute of Virology and Immunology (IVI)MittelhäusernSwitzerland
  2. 2.Centre of Regenerative MedicineUniversity of ManchesterManchesterUK
  3. 3.Centre de Biophysique MoléculaireCNRS UPR4301Orléans cedex 2France
  4. 4.OzBiosciencesParc scientifique de LuminyMarseilleFrance
  5. 5.Department of Vaccinology and Applied MicrobiologyHelmholtz Centre for Infection ResearchBraunschweigGermany

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