Anushi Rajapaksa, Jenny Ho, Aisha Qi, Tri-Hung Nguyen, Michelle Tate, Rob Bischof, David Piedrafita, Michelle McIntosh, Leslie Yeo, Els Meeusen, Ross Coppel and James Friend
Respiratory Research 2014, 15:60 doi:10.1186/1465-9921-15-60
Published: 20 May 2014Abstract (provisional)
Background
Pulmonary-delivered gene therapy promises to mitigate vaccine safety issues and reduce the need for needles and skilled personnel to use them. While plasmid DNA (pDNA) offers a rapid route to vaccine production without side effects or reliance on cold chain storage, its delivery to the lung has proved challenging. Conventional methods, including jet and ultrasonic nebulizers, fail to deliver large biomolecules like pDNA intact due to the shear stresses present during nebulization.
Methods
In vitro structural analysis followed by in vivo protein expression studies served in assessing the integrity of the pDNA subjected to surface acoustic wave (SAW) nebulisation. In vivo immunization trials were then carried out in rats using SAW nebulized pDNA (influenza A, human hemagglutinin H1N1) condensate delivered via intratracheal instillation. Finally, in vivo pulmonary vaccinations using pDNA for influenza was nebulised and delivered via respirator to sheep.
Results
The SAW nebulizer was effective at generating pDNA aerosols with sizes optimal for deep-lung delivery. Successful gene expression was observed in mouse lung epithelial cells, when SAW-nebulized pDNA was delivered to a male Swiss mouse via intratracheal instillation. Effective systemic and mucosal antibody responses was found in rats via post-nebulized, condensed fluid instillation. Significantly, we demonstrated the suitability of the SAW nebulizer to administer unprotected pDNA encoding an influenza A virus surface glycoprotein to respirated sheep via aerosolized inhalation.
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