Related to deliverable No: D.3.1.2
Version 20 Nov. 09
Tool title and potential use
Reporter cell lines for screening pollution-related cellular stress and immune
activation
Novelty and background
Human cell lines were stably transfected with the Luciferase reporter gene under
control of promoters reflecting on cellular stress and immune activation. The
usefulness of this screening tool for the analysis of pollution-related chemicals and
chemical mixtures was verified in the NoMiracle project.
Decription of tool and current state
We have started to produce stable reporter cell lines during the FP5 project
MAAPHRI and could substantially extend the panel of available cell lines within
NOMIRACLE. We and others have sucessfully used our reporter cell lines to
investigate various chemicals, including volatile components and mixtures, as well as
a number of different nanomaterials. These studies have shown that the standardised
reporter gene approach can successfully detect cellular stress and immune activation
which are later verified in more elaborate assays. It is of substantial interest that a
number of cell lines is available since no single assay exists which could give similar
information for screening formats. The follwing Luc-reporter cell lines are at present
available:
In Jurkat:
IL-4, IL-6, IL-8, IL-13, IFN-, TGF-ß, TNF-, NF-B
In A549:
IL-6, IL-8, TNF-, NF-B
In HeLa:
IL-8
In HEK:
Eotaxin-1
The tool is ready to use.
References
1. Holmstrup M, Bindesbol A-M, Oostingh G J, Duschl A, Scheil V, Köhler H,
Loureiro S, Soares A, Ferreira A, Gerhard A, Kienle C, Laskowski R,
Kramarz P, Bayley M, Svendsen C, Spurgeon D (In press). Interactions
between effects of environment chemicals and natural stressors: a review. Sci
Total Environ.
2. Pfaller T, Colognato R, Nelissen I, Flavilli F, Casals E, Ooms D, Leppens H,
Ponti J, Stritzinger R, Puntes V, Boraschi D, Duschl A, Oostingh GJ. (in
press). The suitability of different cellular in vitro immunotoxicity and
genotoxicity methods for the analysis of nanoparticle induced effects.
3.
4.
5.
6.
7.
8.
9.
Nanotoxicology.
Oostingh G J, Wichmann G, Schmittner M, Lehmann I, Duschl A. (2009).
The cytotoxic effects of the organophosphates chlorpyrifos and diazinon
differ from the immunomodulating effects. J. Immunotoxicol. 6: 136-145
Herzog E, Byrne H J, Davoren M, Casey A, Schmittner M, Duschl A,
Oostingh, G J (2009). Dispersion medium modulates oxidative stress
response of human lung epithelial cells upon exposure to carbon nanomaterial
samples. Toxicology and Applied Pharmacology 236: 276-81
Pfaller T, Puntes V, Casals E, Duschl A, Oostingh G J (2009). In vitro
investigation of immunomodulatory effects caused by engineered inorganic
nanoparticles – the impact of experimental design and cell choice.
Nanotoxicology, 3: 46-59
Herzog E, Byrne H J, Casey A, Davoren M, Lenz A-G, Maier K, Duschl A,
Oostingh G J (2009). SWCNT suppress inflammatory mediator responses in
human lung epithelium in vitro. Toxicol. Appl. Pharmacol. 234: 378-90
Röder-Stolinski C, Fischäder G, Oostingh G J, Eder K, Duschl A, Lehmann I
(2008). Chlorobenzene induces the NF-kB and p38 MAP kinase pathways in
lung epithelial cells. Inhal Toxicol, 20: 813-20.
Röder-Stolinski C, Fischäder G, Oostingh G J, Feltens R, Kohse F, von
Bergen M, Mörbt N, Eder K, Duschl A, Lehmann I (2008). Styrene induces
an inflammatory response in human lung epithelial cells via oxidative stress
and NF-kB activation. Toxicol Appl Pharmacol, 23: 241-7. Epub Apr 29.
doi:10.1016/j.taap.2008.04.010
Oostingh G J, Schmittner M, Ehart A K, Tischler U, Duschl A (2008). A
high-throughput screening method based on stably transformed human cells
was used to determine the immunotoxic effects of fluoranthene. Toxicol in
Vitro, 22: 1301-1310.
Responsible scientists:
Albert Duschl, University of Salzburg, Department of Molecular Biology
albert.duschl@sbg.ac.at
Gertie Janneke Oostingh, University of Salzburg, Department of Molecular Biology
geja.oostingh@sbg.ac.at