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Permanent link (DOI): https://doi.org/10.7939/R3W07K

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DENDRITIC CELL-TARGETED NANOPARTICLES FOR THE DELIVERY OF DNA AND PROTEIN VACCINES Open Access

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Other title
Subject/Keyword
Vaccine
Nanoparticle
Dendritic cell targeting
Type of item
Thesis
Degree grantor
University of Alberta
Author or creator
Raghuwanshi,Dharmendra
Supervisor and department
Kamaljit Kaur (Faculty of Pharmacy and Pharmaceutical Sciences)
Examining committee member and department
Michael R. Doschak(Faculty of Pharmacy and Pharmaceutical Sciences)
Afsaneh Lavasanifar (Faculty of Pharmacy and Pharmaceutical Sciences)
Glen S. Kwon (School of Pharmacy, University of Wisconsin-Madison)
Hasan Uludağ (Chemical and Materials Engineering)
Carlos A. Velázquez (Faculty of Pharmacy and Pharmaceutical Sciences)
Department
Faculty of Pharmacy and Pharmaceutical Sciences
Specialization
Pharmaceutical Sciences
Date accepted
2012-08-29T16:07:42Z
Graduation date
2012-11
Degree
Doctor of Philosophy
Degree level
Doctoral
Abstract
Dendritic cells (DCs) play a central role in shaping antigen-specific immune response. Antibody-mediated antigen targeting to DC-specific surface receptors is a promising approach to enhance vaccine efficacy. The objective of this thesis was to develop DC-targeted nanoparticulate formulations for the delivery of DNA and protein antigen using a novel strategy. The approach involved use of a two-component DC targeted delivery system for enhanced immune response. One component consisting of a recombinant bifunctional fusion protein (bfFp) was used for DC targeting, whereas, the other component made of biotinylated nanoparticles encapsulated antigen. For DNA vaccines, two strategies were adapted. In the first strategy, bfFp functionalized biotinylated chitosan nanoparticles containing DNA-encoding for nucleocapsid (N) of severe acute respiratory syndrome coronavirus (SARS-CoV) or hemagglutinin (HA) of avian influenza virus was used for nasal delivery. Immune response studies in mice showed that intranasal administration of targeted formulation along with DC maturation stimuli (anti-CD40 mAb) enhanced magnitude of mucosal, humoral and cellular immune responses. In the second strategy, a DNA (pDECN) vaccine encoding a fusion protein comprised of SARS CoV N antigen and anti-DEC-205 scFv was constructed. In vitro studies showed that expressed protein was able to bind with DCs. Vaccination of mice with pDECN-laoded chitosan nanoparticles induced significantly higher IgG and cytokine (IFN-γ and IL-2) response relative to SARS CoV N DNA. Coadministration of anti-CD40 antibody further improved efficacy of nanoencapsulated DNA formulations. For the delivery of a model antigen ovalbumin (OVA), biotinylated poly(D,L-lactic-co-glycolic acid) (PLGA) nanoparticles were formulated using biotin-PEG-PLGA polymer and were decorated with bfFp. In vitro uptake studies revealed one-fold higher uptake of targeted nanoparticles compared to non-targeted NPs. In vivo studies show targeted NPs in conjunction with anti-CD40 mAb enhanced OVA-specific IgG and IgG subclass responses. Splenocytes of these mice secreted significantly higher levels of IFN-γ and IL-2, indicating Th1 response. In conclusion, these results demonstrate that bfFp based DC targeting is a versatile approach and vaccine efficacy can be enhanced via non-invasive DC targeting. The two-component DC targeting approach can serve as a viable alternative to conventional antibody-targeted vaccines that also precludes any post-formulation modification of the antigen-loaded NPs.
Language
English
DOI
doi:10.7939/R3W07K
Rights
Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of these terms. The author reserves all other publication and other rights in association with the copyright in the thesis and, except as herein before provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission.
Citation for previous publication
Raghuwanshi D, Mishra V, Das D, Kaur K, Suresh MR. Dendritic cell targeted chitosan nanoparticles for nasal DNA immunization against SARS CoV nucleocapsid protein. Molecular Pharmaceutics 2012;9:946-56

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