Boisen, Anja

Boyd, B.

De Rossi, C.

Gordon, S.

Lehr, C-M.

Nielsen, Line Hagner

Rades, T.

A triple co-culture cell model of Caco-2 cells, dendritic cells and macrophages (Figure 1) has previously been developed for studying intestinal permeability in a state of inflammation [1],[2]. The aim of this study was to investigate the applicability of this cell model for testing the immunostimulatory ability of particulate vaccine formulations designed for oral delivery. Levels of cytokine production in response to vaccine administration were measured following particulate vaccine administration, as an indication of dendritic cell and macrophage activation. Precursors of cubosomes containing the model antigen ovalbumin was spray dried to obtain a particulate vaccine model system for testing in the cell model. The precursors were shown to form cubosomes when dispersed in aqueous medium, and was therefore used as the vaccine formulation for testing on the co-cultures. After 11 days, the TEER values of the co-cultures were found to be 860-1340 Ω∙cm2; the formulations were incubated with the co-cultures at this time point. From confocal microscopy images, it was observed that the THP-1 cells (macrophages) migrated into the overlying Caco-2 cell monolayer when the co-cultures were incubated with particle formulations. This was not the case when incubating with ovalbumin solution or blank. The ELISA screening assay showed production of a wide range of cytokines following culture incubation with cubosomes (with and without ovalbumin) and LPS solutions, indicative of a stimulatory effect; this was not observed with ovalbumin and blank solution. An example of the results is shown in Figure 2 for IL-17A. An established co-culture of Caco-2, THP-1 and MUTZ-3 cells showed promise as an in vitro model for testing of oral vaccine formulations. Mobility of co-culture immune cells as well as cytokine production observed following treatment with spray dried cubosomes as a particulate vaccine formulation will be further investigated. <br/

Online Research Database In Technology

                       General rights Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights.  • Users may download and print one copy of any publication from the public portal for the purpose of private study or research. • You may not further distribute the material or use it for any profit-making activity or commercial gain • You may freely distribute the URL identifying the publication in the public portal   If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim.     Downloaded from orbit.dtu.dk on: Nov 09, 2017Triple co-culture cell model as an in vitro model for oral particulate vaccine systemsNielsen, Line Hagner; De Rossi, C.; Lehr, C-M.; Rades, T.; Boyd, B.; Boisen, Anja; Gordon, S.Publication date:2016Document VersionPublisher's PDF, also known as Version of recordLink back to DTU OrbitCitation (APA):Nielsen, L. H., De Rossi, C., Lehr, C-M., Rades, T., Boyd, B., Boisen, A., & Gordon, S. (2016). Triple co-culturecell model as an in vitro model for oral particulate vaccine systems. Poster session presented at BioBarriers2016, Saarbrücken, Germany.Please create your abstract using the following template and send the document as pdf-file to  Biobarriers2016@mx.uni-saarland.de Please do not modify the format of this template! Abstract limit is 1 page A4.  NOTE: Please save your abstract file according to the name of the presenting author as a ‘pdf ’ file (Author A B.pdf).   Triple co-culture cell model as an in vitro model for oral particulate vaccine systems Hagner Nielsen L.1, De Rossi C.2, Lehr C-M.2,3, Rades T.4, Boyd B.5, Boisen A.1, Gordon S.2 1Department of Micro and Nanotechnology, Technical University of Denmark, Kgs. Lyngby, Denmark  2Department of Drug Delivery, Helmholtz Institute for Pharmaceutical Research Saarland, Helmholtz Centre for Infection Research, Saarbrücken, Germany 3Department of Pharmacy, Biopharmacy and Pharmaceutical Technology, Saarland University, Saarbrücken, Germany 4Department of Pharmacy, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark  5Monash Institute of Pharmaceutical Sciences, Monash University, Melbourne, Australia  A triple co-culture cell model of Caco-2 cells, dendritic cells and macrophages (Figure 1) has previously been developed for studying intestinal permeability in a state of inflammation [1],[2].  The aim of this study was to investigate the applicability of this cell model for testing the immunostimulatory ability of particulate vaccine formulations designed for oral delivery. Levels of cytokine production in response to vaccine administration were measured following particulate vaccine administration, as an indication of dendritic cell and macrophage activation. Precursors of cubosomes containing the model antigen ovalbumin was spray dried to obtain a particulate vaccine model system for testing in the cell model. The precursors were shown to form cubosomes when dispersed in aqueous medium, and was therefore used as the vaccine formulation for testing on the co-cultures.  After 11 days, the TEER values of the co-cultures were found to be 860-1340 Ω∙cm2; the formulations were incubated with the co-cultures at this time point. From confocal microscopy images, it was observed that the THP-1 cells (macrophages) migrated into the overlying Caco-2 cell monolayer when the co-cultures were incubated with particle formulations. This was not the case when incubating with ovalbumin solution or blank. The ELISA screening assay showed production of a wide range of cytokines following culture incubation with cubosomes (with and without ovalbumin) and LPS solutions, indicative of a stimulatory effect; this was not observed with ovalbumin and blank solution. An example of the results is shown in Figure 2 for IL-17A.    An established co-culture of Caco-2, THP-1 and MUTZ-3 cells showed promise as an in vitro model for testing of oral vaccine formulations. Mobility of co-culture immune cells as well as cytokine production observed following treatment with spray dried cubosomes as a particulate vaccine formulation will be further investigated.   Acknowledgments: The authors would like to acknowledge the Danish Research Council for Technology and Production (FTP), Project DFF - 4004-00120B for financial support. Moreover, the Danish National Research Foundation, project: DNRD122 is acknowledged.  References: 1. Leonard, F., Collnot, EM., Lehr, C-M.: Mol Pharm. 2010, 7: 2103-2119. 2. Susewind, J. et al.: Nanotoxicology, 2015, 4: 1-10.   

Triple co-culture cell model as an in vitro model for oral particulate vaccine systems

http://orbit.dtu.dk/ws/files/125196040/BB2016_cell_model.pdf

Triple co-culture cell model as an in vitro model for oral particulate vaccine systems

Abstract

Similar works

Full text

Available Versions

Online Research Database In Technology