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Cationic lipid/pDNA complex formation as potential generic method to generate specific IRF pathway stimulators.

Simone Putzke1, Elisabeth Feldhues2, Iris Heep2

  • 1Department of Pharmaceutics, Institute of Pharmacy, University of Bonn, 53121 Bonn, Germany; Bayer Animal Health GmbH, 40789 Monheim am Rhein, Germany.

European Journal of Pharmaceutics and Biopharmaceutics : Official Journal of Arbeitsgemeinschaft Fur Pharmazeutische Verfahrenstechnik E.V
|August 18, 2020
PubMed
Summary

Cationic liposome-CpG DNA complexes selectively activate the interferon response factor (IRF) pathway, leading to significant cytokine secretion. These lipoplexes enhance DNA uptake and bypass Toll-like receptor 9 (TLR9) and NF-κB activation.

Keywords:
Cationic lipid/pDNA complexesDendritic cellsIRF pathwayInnate immunityLipoplexesMacrophagesNF-κB pathwayType I interferon

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Area of Science:

  • Immunology
  • Nanotechnology
  • Molecular Biology

Background:

  • Cationic liposome-CpG DNA complexes (lipoplexes) activate innate immunity via Toll-like receptor 9 (TLR9) and the nuclear factor kappa B (NF-κB) pathway.
  • Emerging evidence suggests lipoplexes also engage cytosolic DNA sensors, stimulating the interferon response factor (IRF) pathway.

Purpose of the Study:

  • To formulate and characterize various lipoplexes using different cationic lipids and CpG plasmids.
  • To investigate the cellular uptake and immune signaling pathway activation induced by these lipoplexes.

Main Methods:

  • Formulation of lipoplexes using DOTAP, DOTMA, DDA cationic lipids and varying CpG plasmids.
  • Characterization of lipoplex physicochemical properties (size, charge, loading).
  • Assessment of cellular lipid and DNA uptake in macrophages and dendritic cells.
  • Analysis of NF-κB and IRF pathway activation using reporter cell lines.
  • Measurement of interferon-alpha and -beta secretion.

Main Results:

  • Lipoplexes showed consistent physicochemical properties regardless of lipid or plasmid composition.
  • Significant increase in cellular lipid and DNA uptake with lipoplexes compared to naked DNA.
  • Massive activation of the IRF pathway (up to 46-fold) with minimal NF-κB activation.
  • DOTAP and DOTMA lipoplexes induced substantial interferon-alpha and -beta secretion.

Conclusions:

  • CpG plasmid DNA lipoplexes selectively activate the IRF pathway, bypassing TLR9 and NF-κB.
  • Lipoplexes facilitate efficient DNA delivery into the cytosol via vesicular compartments.
  • These findings highlight the potential of tailored lipoplexes for targeted immune stimulation.