"Fluorescent Cell Chip" for immunotoxicity testing: development of the c-fos expression reporter cell lines.
2005 (English)In: Toxicology and Applied Pharmacology, ISSN 0041-008X, E-ISSN 1096-0333, Vol. 207, no 2 Suppl, 133-41 p.Article in journal (Refereed) Published
The Fluorescent Cell Chip for in vitro immunotoxicity testing employs cell lines derived from lymphocytes, mast cells, and monocytes-macrophages transfected with various EGFP cytokine reporter gene constructs. While cytokine expression is a valid endpoint for in vitro immunotoxicity screening, additional marker for the immediate-early response gene expression level could be of interest for further development and refinement of the Fluorescent Cell Chip. We have used BW.5147.3 murine thymoma transfected with c-fos reporter constructs to obtain reporter cell lines expressing ECFP under the control of murine c-fos promoter. These cells upon serum withdrawal and readdition and incubation with heavy metal compounds showed paralleled induction of c-Fos expression as evidenced by Real-Time PCR and ECFP fluorescence as evidenced by computer-supported fluorescence microscopy. In conclusion, we developed fluorescent reporter cell lines that could be employed in a simple and time-efficient screening assay for possible action of chemicals on c-Fos expression in lymphocytes. The evaluation of usefulness of these cells for the Fluorescent Cell Chip-based detection of immunotoxicity will require additional testing with a larger number of chemicals.
Place, publisher, year, edition, pages
2005. Vol. 207, no 2 Suppl, 133-41 p.
IdentifiersURN: urn:nbn:se:uu:diva-161183DOI: 10.1016/j.taap.2005.01.052PubMedID: 16023160OAI: oai:DiVA.org:uu-161183DiVA: diva2:455150