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Optimizing Adenoviral Transduction of Endothelial Cells under Flow Conditions

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ABSTRACT

Purpose

To target adenoviral vectors to cells of the vasculature and shielding vectors from inactivation by the immune system.

Methods

Complexes of reporter gene expressing adenoviral vectors with positively charged magnetic nanoparticles were formed by electrostatic interaction in presence or absence of additional negatively charged poly(ethylene glycol)-based polymer. Transduction of HUVEC was analyzed in vitro under flow. Protection from inactivation by the immune system was analyzed by pre-incubation of AdV and complexes with neutralizing antibodies and subsequent reporter protein analysis of infected cells.

Results

Physical association of AdV with MNP and polymers was demonstrated by radioactive labelling of components and co-sedimentation in a magnetic field. Ad-MNP+/−polymer resulted in efficient transduction of HUVEC, depending on MOI and flow rate in presence of magnetic field, whereas no transduction was observed without complex formation with MNP or in absence of magnetic field. Association with MNP did result in protection from neutralizing antibodies, with slightly increased protection provided by the polymer.

Conclusions

Complex formation of AdV with MNP is a viable means for targeting of vectors to areas of magnetic field gradient. Additional coating with polymer might proof useful in protection from inactivation by the immune system.

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Abbreviations

AdV:

adenoviral vectors

CMV:

cytomegalovirus

eGFP:

enhanced green fluorescent protein

HUVEC:

human umbilical cord venous endothelial cells

Luc:

firefly luciferase

MNP:

magnetic nanoparticles

MOI:

multiplicity of infection

PBS:

phosphate buffered saline

PEG:

poly(ethylene glycol)

PEI:

poly(ethylene imine)

PFU:

plaque forming units

PM:

polymer P6YE5C

VP:

virus particles

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Acknowledgments & Disclosures

This work was supported by the German Research Foundation, through DFG Research Unit FOR917 Nanoguide (AN 333/1-1 and Projects PL 281/3-1). We thank Katja Dumler for excellent technical assistance.

C.P. is a co-founder of OZ Biosciences, SA, Marseille France. OZBiosciences commercializes reagents for magnetofection.

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Authors and Affiliations

  1. Institute of Experimental Oncology and Therapy Research, Klinikum rechts der Isar der Technischen Universität München, Munich, Germany

    Martina Anton, Anja Wolf, Olga Mykhaylyk, Christian Koch, Bernd Gansbacher & Christian Plank

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  1. Martina Anton
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  2. Anja Wolf
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  3. Olga Mykhaylyk
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Correspondence to Martina Anton.

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Anton, M., Wolf, A., Mykhaylyk, O. et al. Optimizing Adenoviral Transduction of Endothelial Cells under Flow Conditions. Pharm Res 29, 1219–1231 (2012). https://doi.org/10.1007/s11095-011-0631-2

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  • DOI: https://doi.org/10.1007/s11095-011-0631-2

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