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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
VerfasserStoehr, Linda C. ; Endes, Carola ; Radauer-Preiml, Isabella ; Boyles, Matthew S. P. ; Casals, Eudald ; Balog, Sandor ; Pesch, Markus ; Petri-Fink, Alke ; Rothen-Rutishauser, Barbara ; Himly, Martin ; Clift, Martin J. D. ; Duschl, Albert
Erschienen in
Particle and Fibre Toxicology, London : BioMed Central, 2015, Jg. 12, H. 29, S. 1-12
SpracheEnglisch
DokumenttypAufsatz in einer Zeitschrift
Schlagwörter (EN)A549 cells / Interleukin-8 / Air-liquid interface / Submerged cultures / Acute pulmonary (pro-)inflammatory effects
ISSN1743-8977
URNurn:nbn:at:at-ubs:3-3986 Persistent Identifier (URN)
DOI10.1186/s12989-015-0104-6 
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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 [1.13 mb]
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BACKGROUND: 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. METHODS: 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). RESULTS: 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. CONCLUSIONS: 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.

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