Combined use of radioimagers and radioactive 3'OH DNA nick end labelling to quantify apoptosis in cell lines and tissue sections: applications to virus-induced apoptosis.
DNA fragmentation is a key feature of the degradation phase of apoptosis. In this work we have developed an assay, based on radioimager (beta-IMAGER and micro-IMAGER) quantification of radioactive nick end labelling (RANEL), which is quantitative, rapid and sensitive to study in vitro and in vivo induced apoptosis. To establish the technique, in vitro apoptosis of T cell lines was induced by stimulation of the Fas receptor; cells were labelled using TdT-mediated [alpha-33P] dCTP nick end labelling, after which then radioactivity was quantified using a beta-IMAGER. We have also shown that the RANEL method can be applied to the quantification and visualisation, by micro-IMAGER analysis, of liver tissue sections from mouse Fas-induced fulminant hepatitis or from Dengue-1 virus infected individuals. Finally, this system has also been used to detect apoptosis induced by rabies virus in Jurkat T cells. These data have established a large field of application for the RANEL assay.
Jacotot
E
E
Laboratoire de Technologie Cellulaire, Département des Biotechnologies, Institut Pasteur, 25-28 rue Dr. Roux, 75724 Paris, France.
Cardona
A
A
Rebouillat
D
D
Terradillos
O
O
Marianneau
P
P
Thoulouze
M I
MI
Lafon
M
M
Deubel
V
V
Edelman
L
L