Fate of TLR-1/TLR-2 agonist functionalised pDNA nanoparticles upon deposition at the human bronchial epithelium in vitro

Journal of Nanobiotechnology

Research

Peter Gehr¹  David Olivier Raemy²  Barbara Rothen-Rutishauser²  Gerrit Borchard³  Simon Heuking⁴ 

[1] Institute of Anatomy, University of Bern, Bern, Switzerland;Institute of Anatomy, University of Bern, Bern, Switzerland;Adolphe Merkle Institute, University of Fribourg, Fribourg, Switzerland;School of Pharmaceutical Sciences Geneva-Lausanne (EPGL), University of Geneva, University of Lausanne, Geneva, Switzerland;School of Pharmaceutical Sciences Geneva-Lausanne (EPGL), University of Geneva, University of Lausanne, Geneva, Switzerland;Institute of Anatomy, University of Bern, Bern, Switzerland;Vaccine Formulation Laboratory, Department of Biochemistry, University of Lausanne, Lausanne, Switzerland;

关键词: Triple cell co-culture;    Dendritic cells;    Macrophages;    Bronchial epithelium;    Toll-like receptor agonist;    Adjuvant;    Chitosan DNA nanoparticles;   

DOI  :  10.1186/1477-3155-11-29

 received in 2013-06-17, accepted in 2013-08-16,  发布年份 2013

来源: Springer

【 摘 要 】

BackgroundPlasmid DNA vaccination is a promising approach, but studies in non-human primates and humans failed to achieve protective immunity. To optimise this technology further with focus on pulmonary administration, we developed and evaluated an adjuvant-equipped DNA carrier system based on the biopolymer chitosan. In more detail, the uptake and accompanying immune response of adjuvant Pam3Cys (Toll-like receptor-1/2 agonist) decorated chitosan DNA nanoparticles (NP) were explored by using a three-dimensional (3D) cell culture model of the human epithelial barrier. Pam3Cys functionalised and non-functionalised chitosan DNA NP were sprayed by a microsprayer onto the surface of 3D cell cultures and uptake of NP by epithelial and immune cells (blood monocyte-derived dendritic cells (MDDC) and macrophages (MDM)) was visualised by confocal laser scanning microscopy. In addition, immune activation by TLR pathway was monitored by analysis of interleukin-8 and tumor necrosis factor-α secretions (ELISA).ResultsAt first, a high uptake rate into antigen-presenting cells (MDDC: 16-17%; MDM: 68–75%) was obtained. Although no significant difference in uptake patterns was observed for Pam3Cys adjuvant functionalised and non-functionalised DNA NP, ELISA of interleukin-8 and tumor necrosis factor-α demonstrated clearly that Pam3Cys functionalisation elicited an overall higher immune response with the ranking of Pam3Cys chitosan DNA NP > chitosan DNA NP = DNA unloaded chitosan NP > control (culture medium).ConclusionsChitosan-based DNA delivery enables uptake into abluminal MDDC, which are the most immune competent cells in the human lung for the induction of antigen-specific immunity. In addition, Pam3Cys adjuvant functionalisation of chitosan DNA NP enhances significantly an environment favoring recruitment of immune cells together with a Th1 associated (cellular) immune response due to elevated IL-8 and TNF-α levels. The latter renders this DNA delivery approach attractive for potential DNA vaccination against intracellular pathogens in the lung (e.g., Mycobacterium tuberculosis or influenza virus).

【 授权许可】

Unknown   
© Heuking et al.; licensee BioMed Central Ltd. 2013. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.