"Fluorescent Cell Chip" for immunotoxicity testing: development of the c-fos expression reporter cell lines.
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.
Trzaska
Dominika
D
International Institute of Molecular and Cell Biology, Warsaw, Poland.
Zembek
Patrycja
P
Olszewski
Maciej
M
Adamczewska
Violetta
V
Ullerås
Erik
E
Dastych
Jarosław
J
D000826:Animal Testing Alternatives; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers; D017930:Genes, Reporter; D016762:Genes, fos; D007107:Immune System / Q000187:drug effects*; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017931:DNA Primers