Assessment of a panel of interleukin-8 reporter lung epithelial cell lines to monitor the pro-inflammatory response following zinc oxide nanoparticle exposure under different cell culture conditions
Stoehr, Linda C. (Paris-Lodron-Universität Salzburg. Department of Molecular Biology)
Endes, Carola (Universität Freiburg. BioNanomaterials, Adolphe Merkle Institute)
Radauer-Preiml, Isabella (Paris-Lodron-Universität Salzburg. Department of Molecular Biology)
Boyles, Matthew S. P. (Paris-Lodron-Universität Salzburg. Department of Molecular Biology)
Casals, Eudald (Institut Català de Nanociència i Nanotecnologia)
Balog, Sandor (Universität Freiburg. Soft Matter Scattering)
Pesch, Markus (Grimm Aerosol Technik GmbH & Co. KG, Ainring)
Petri-Fink, Alke (Universität Freiburg. BioNanomaterials, Adolphe Merkle Institute)
Rothen-Rutishauser, Barbara (Universität Freiburg. BioNanomaterials, Adolphe Merkle Institute)
Himly, Martin (Paris-Lodron-Universität Salzburg. Department of Molecular Biology)
Clift, Martin J. D. (Universität Freiburg. BioNanomaterials, Adolphe Merkle Institute)
Duschl, Albert (Paris-Lodron-Universität Salzburg. Department of Molecular Biology)

Data:	2015
Resum:	Stably transfected lung epithelial reporter cell lines pose an advantageous alternative to replace complex experimental techniques to monitor the pro-inflammatory response following nanoparticle (NP) exposure. Previously, reporter cell lines have been used under submerged culture conditions, however, their potential usefulness in combination with air-liquid interface (ALI) exposures is currently unknown. Therefore, the aim of the present study was to compare a panel of interleukin-8 promoter (pIL8)-reporter cell lines (i. e. green or red fluorescent protein (GFP, RFP), and luciferase (Luc)), originating from A549 lung epithelial type II-like cells cells, following NPs exposure under both submerged and ALI conditions. All cell lines were exposed to zinc oxide (ZnO) NPs at 0. 6 and 6. 2 μg/cm 2 for 3 and 16 hours under both submerged and ALI conditions. Following physicochemical characterization, the cytotoxic profile of the ZnO-NPs was determined for each exposure scenario. Expression of IL-8 from all cell types was analyzed at the promoter level and compared to the mRNA (qRT-PCR) and protein level (ELISA). In summary, each reporter cell line detected acute pro-inflammatory effects following ZnO exposure under each condition tested. The pIL8-Luc cell line was the most sensitive in terms of reporter signal strength and onset velocity following TNF-α treatment. Both pIL8-GFP and pIL8-RFP also showed a marked signal induction in response to TNF-α, although only after 16 hrs. In terms of ZnO-NP-induced cytotoxicity pIL8-RFP cells were the most affected, whilst the pIL8-Luc were found the least responsive. In conclusion, the use of fluorescence-based reporter cell lines can provide a useful tool in screening the pro-inflammatory response following NP exposure in both submerged and ALI cell cultures. The online version of this article (doi:10. 1186/s12989-015-0104-6) contains supplementary material, which is available to authorized users.
Ajuts:	European Commission 263147 European Commission 264506
Drets:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Llengua:	Anglès
Document:	Article ; recerca ; Versió publicada
Matèria:	A549 cells ; Interleukin-8 ; Air-liquid interface ; Submerged cultures ; Acute pulmonary (pro-)inflammatory effects
Publicat a:	Particle and fibre toxicology, Vol. 12 (September 2015) , art. 29, ISSN 1743-8977

DOI: 10.1186/s12989-015-0104-6
PMID: 26415698

12 p, 1.1 MB

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