Fluorescent Cell Chip a new in vitro approach for immunotoxicity screening.
The Fluorescent Cell Chip (FCC) has been developed specifically for immunotoxicity screening of chemical compounds. This in vitro test is based on a panel of genetically modified reporter cell lines that regulate the expression of fluorescent protein in the same way as they regulate expression of cytokines. Thus, changes in fluorescence intensity represent changes in cytokine expression. Consequently, this technique conforms to efficiency expected from high throughput screening assay. In a pre-validation effort we analyzed 46 compounds. The experimental protocol employed five reporter cell lines derived from murine EL-4 T cells. Reporter cells were exposed to tested chemicals on a 96 well plate and analyzed for EGFP-mediated fluorescence using automated flow cytometric assay. Tested compounds reproducibly generated compound-specific patterns of changes in fluorescence that allows for the hierarchical clustering of their expected activities based on pattern similarity analysis. Resultant classification revealed correlation with available in vivo immunotoxicity data. In conclusion, FCC is a new promising approach for in vitro screening of chemicals for their immunotoxicity.
Wagner
Waldemar
W
Centre for Medical Biology of Polish Academy of Sciences, Lodowa 106, 93-232 Łódź, Poland.
Walczak-Drzewiecka
Aurelia
A
Slusarczyk
Aleksandra
A
Biecek
Przemysław
P
Rychlewski
Leszek
L
Dastych
Jarosław
J
D000818:Animals; D002461:Cell Line, Transformed; D064420:Drug-Related Side Effects and Adverse Reactions; D005434:Flow Cytometry; D005453:Fluorescence; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D007155:Immunologic Factors / Q000633:toxicity*; D007287:Inorganic Chemicals / Q000633:toxicity; D051379:Mice; D009930:Organic Chemicals / Q000633:toxicity; D013601:T-Lymphocytes / Q000276:immunology; D018675:Toxicity Tests / Q000379:methods*
D007155:Immunologic Factors; D007287:Inorganic Chemicals; D009930:Organic Chemicals; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins