We have shown that both uptake and gene expression (transcription of reporter gene) of PLL/DNA polyplexes are dependent on DNA topology. Complexes selleck chemicals containing SC-pDNA were most efficient in associating with the nucleus (polyplex fluorescence overlaid with nuclear stain) as observed by confocal microscopy studies (15% [2.59% RSE] associated with the nucleus in comparison to no nuclear association reported for OC- and linear-pDNA at 1 h). However confocal quantification via fluorescence overlay does not directly

correspond to gene expression, as nuclear uptake of DNA can still be hindered by the presence of nucleases [9]. Complexes containing SC-pDNA displayed significantly higher gene expression (14%) than other topological forms (9.59% and 7.43% for OC- and linear-pDNA polyplexes) (p < 0.05), although expression was modest in comparison to that reported for CHO cells [9]. This may be due to DCs predominately expressing nucleases which restrict

uptake and gene expression. PF2341066 Lack of DC surface marker expression may be explained by low dosage (20 μg) used. This in itself may be considered advantageous in terms of biocompatibility and safe delivery of DNA in vivo [21]. In terms of bio-processing and vaccine production, the application of SC-pDNA is a key pre-requisite. The findings of this study show how pDNA in the SC conformation is more efficient in terms of both uptake and gene expression than OC- and linear-pDNA. Therefore DNA topology does impact on processing and vaccine manufacture. This is in agreement with current regulatory bodies such as the FDA which require second 80% SC content (Guidance for Industry: Considerations for Plasmid DNA Vaccines for Infectious Disease Indications – FDA, 2007) [26]. The authors would like to thank the Engineering and Physical Sciences Research Council (EPSRC) for both the PhD studentship support for Arjun Dhanoya and the sponsorship of the Innovative Manufacturing Research Council (IMRC)

for Bioprocessing at UCL. We also thank Dr. Nicola Hardwick for advice and technical support. “
“During the A/H1N1 2009–2010 pandemic, up to 33.0% of influenza cases, 32.0% of hospitalizations and 10.0% of deaths due to influenza in the US were reported for individuals younger than 18 years of age [1] and [2]. In Europe, data from the European Influenza Surveillance Network showed that the highest rates of infection were in school-age children, most cases being mild in severity [3]. When mortality data where compared with those from previous years, excess mortality was observed only in children 5–14 years old [3]. Results from serosurveys showed pre-existing immunity against H1N1/2009 in older persons, with cross-reactive antibodies detected pre-vaccination in 29.8% of people ≥70 years old [4] and 34% in people ≥60 years old [5].