Enhanced delivery and expression of a nanoencapsulated DNA vaccine vector for respiratory syncytial virus

This study evaluated the efficiency of chitosan-encapsulated DNA-based respiratory syncytial virus (RSV) vaccine. Antigenic regions of RSV F, M2, and G genes were cloned into the human cytomegalovirus promoter-based constitutive expression vector, resulting in a DNA vaccine vector named DR-FM2G. This vector was used to formulate DNA-chitosan nanoparticles (DCNPs) using a complex coacervation process that yielded an encapsulation efficiency of 94.7%. The DCNP sizes ranged from 80 to 150 nm with uniform size distribution and spherical shape. DNA release was between 50% and 60% when DCNPs were incubated with similar gastrointestinal fluid (pH 2), whereas 21% to 25% of DNA was released from DCNPs in 30 minutes at pH 10. Differential scanning calorimetry showed DCNPs to be more stable than naked DNA or chitosan, offering protection from DNA degradation by nucleases. DCNPs were not toxic to cells when used at concentrations <= 400 mu g/mL. Immunohistochemical and real-time polymerase chain reaction results showed a higher level of RSV protein expression in mouse tissues given when DCNPs were injected intravenously as compared with naked DNA. From the Clinical Editor: This study evaluated the efficiency of chitosan-encapsulated DNA-based respiratory syncytial virus (RSV) vaccine, showing a higher level of RSV protein expression in mouse tissues given when DCNPs were injected intravenously as compared with naked DNA. (C) 2009 Elsevier Inc. All rights reserved.