"Fluorescent Cell Chip" for immunotoxicity testing: development of the c-fos expression reporter cell lines

Toxicol Appl Pharmacol. 2005 Sep 1;207(2 Suppl):133-41. doi: 10.1016/j.taap.2005.01.052.

Abstract

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.

Publication types

  • Review

MeSH terms

  • Animal Testing Alternatives
  • Animals
  • Base Sequence
  • Cell Line
  • DNA Primers
  • Genes, Reporter*
  • Genes, fos*
  • Immune System / drug effects*
  • Mice
  • Microscopy, Fluorescence / methods*
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • DNA Primers