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Virology Journal
Published in: Virology Journal 1/2009

Open Access 01-12-2009 | Methodology

Analysis of adenoviral attachment to human platelets

Authors: Nilly Shimony, Gregory Elkin, Dror Kolodkin-Gal, Lina Krasny, Simcha Urieli-Shoval, Yosef S Haviv

Published in: Virology Journal | Issue 1/2009

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Abstract

Background

Systemic adenoviral (Ad) vector administration is associated with thrombocytopenia. Recently, Ad interaction with mouse platelets emerged as a key player determining liver uptake and platelet clearance. However, whether Ad can activate platelets is controversial. Thus, in vitro analysis of Ad attachment to platelets is of interest.

Methods

We developed a direct flow cytometry assay to specifically detect Ad particles adherent to human platelets. The method was pre-validated in nucleated cells. Blocking assays were employed to specifically inhibit Ad attachment to platelets. Platelet activation was analyzed using annexin v flow cytometry.

Results

We found in vitro that Ad binding to human platelets is synergistically enhanced by the combination of platelet activation by thrombin and MnCl2 supplementation. Of note, Ad binding could activate human platelets. Platelets bound Ad displaying an RGD ligand in the fiber knob more efficiently than unmodified Ad. In contrast to a previous report, CAR expression was not detected on human platelets. Integrins appear to mediate Ad binding to platelets, at least partially. Finally, αIIbβ3-deficient platelets from a patient with Glanzmann thrombasthenia could bind Ad 5-fold more efficiently than normal platelets.

Conclusion

The flow cytometry methodology developed herein allows the quantitative measurement of Ad attachment to platelets and may provide a useful in vitro approach to investigate Ad interaction with platelets.
Appendix
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Metadata
Title
Analysis of adenoviral attachment to human platelets
Authors
Nilly Shimony
Gregory Elkin
Dror Kolodkin-Gal
Lina Krasny
Simcha Urieli-Shoval
Yosef S Haviv
Publication date
01-12-2009
Publisher
BioMed Central
Published in
Virology Journal / Issue 1/2009
Electronic ISSN: 1743-422X
DOI
https://doi.org/10.1186/1743-422X-6-25

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