![]() ![]() Plasmid DNA was quantified in blood samples and mRNA expression for IFN-g in tissue samples. Samples were analysed using quantitative real-time PCR. Peripheral blood and biopsies from the injection site were taken at 13 time points until day 14 post injection (p.i.). Seven grey horses bearing melanoma were injected intratumourally with 250 µg naked plasmid DNA coding for IL-12. The expression of induced interferon gamma (IFN-g) was evaluated in order to determine the pharmacodynamic properties of in vivo gene transduction. Whole blood pharmacokinetics of intratumourally injected naked plasmid DNA coding for equine Interleukin 12 (IL-12) was assessed as a means of in vivo gene transfer in the treatment of melanoma in grey horses. Müller, J-M V Wissemann, J Meli, M L Dasen, G Lutz, H Heinzerling, L Feige, K In vivo induction of interferon gamma expression in grey horses with metastatic melanoma resulting from direct injection of plasmid DNA coding for equine interleukin 12. The study signifies the potential of chitosan nanoparticles as DNA vaccine carrier and adjuvant for effective immunization through non-invasive nasal route. Chitosan nanoparticles thus produced humoral (both systemic and mucosal) and cellular immune responses upon nasal administration. Similarly, cellular responses (cytokine levels) were poor in case of alum adsorbed HBsAg. However, intramuscular administration of naked DNA and alum adsorbed HBsAg did not elicit sIgA titre in mucosal secretions that was induced by nasal immunization with chitosan nanoparticles. ![]() Nasal administration of nanoparticles resulted in serum anti-HBsAg titre that was less compared to that elicited by naked DNA and alum adsorbed HBsAg, but the mice were seroprotective within 2 weeks and the immunoglobulin level was above the clinically protective level. Prepared nanoparticles were characterized for size, shape, surface charge, plasmid loading and ability of nanoparticles to protect DNA against nuclease digestion and for their transfection efficacy. Chitosan p DNA nanoparticles were prepared using a complex coacervation process. This work investigates the preparation and in vivo efficacy of plasmid DNA loaded chitosan nanoparticles for nasal mucosal immunization against hepatitis B. Khatri, Kapil Goyal, Amit K Gupta, Prem N Mishra, Neeraj Vyas, Suresh P Plasmid DNA loaded chitosan nanoparticles for nasal mucosal immunization against hepatitis B. These in vivo findings demonstrate the superior stability of PEGylated polyplexes in physiological milieu and provide important insight on how cationic polymers could be optimized for DNA vaccine delivery. By day 4, co-localization of polyplexes with APCs was observed at the injection site regardless of polymer structure, whereas small amounts of polyplexes were found in the draining lymph nodes. PEGylated polyplexes also distributed more broadly among dermal fibroblasts and allowed greater interaction with antigen-presenting cells (APCs) (dendritic cells and macrophages) starting at around 24 h post-injection. The PEGylated polyplexes had a significantly wider distribution in the tissue than the nonPEGylated polyplexes. However, naked plasmid level dropped to below detection limit after 24 h, whereas polyplexes persisted for up to 2 weeks. Live animal imaging revealed that naked plasmid dispersed quickly in the skin of mice after injection and had a wider distribution than any of the three types of polyplexes. At day 4, however, the polyplexes appeared to result in more transfected skin cells than naked plasmid. Comparable numbers of luciferase expressing cells were observed in the skin of mice receiving naked plasmid or polyplexes one day after transfection. We analyzed transgene expression (luciferase) and the local tissue distribution of the labeled plasmid at the injection site at various time points (from hours to days). To better understand the details of DNA vaccine delivery in vivo, we prepared polymer/ DNA complexes using three structurally distinct cationic polymers and fluorescently labeled plasmid DNA and injected them intradermally into mice. Noelle Zhong, Xiao Panus, David Han, Wenqing Ji, Weihang Wang, ChunĭNA vaccination using cationic polymers as carriers has the potential to be a very powerful method of immunotherapy, but typical immune responses generated have been less than robust. Transgene expression and local tissue distribution of naked and polymer-condensed plasmid DNA after intradermal administration in mice ![]()
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