An anti-Bcl-2 antibody prevents 2-deoxy-D-ribose-induced apoptosis in the IPLB-LdFB insect cell line.
Confocal microscopy reveals that the anti-Bcl-2 antibody (pAb) is able to diffuse across the plasma membrane of the fat body cell line IPLB-LdFB from the insect Lymantria dispar, demonstrating the presence of Bcl-2-like molecules in the cytoplasm. Immunoperoxidase procedure confirms the cellular localization. Furthermore, an immunoprecipitation corresponding to a molecular weight of 29 KDa is observed with western blot analysis using the anti-Bcl-2 pAb. Cytofluorimetric experiments show that anti-Bcl-2 pAb counteracts 2-deoxy-D-ribose-induced apoptosis and provokes morphological changes in the insect cell line, i. e. a reduction in cell size, the disappearance of the vacuola and changes in shape. At the same time, the antibody provokes mitochondrial membrane depolarization, and N-acetyl-L-cysteine is unable to reconstitute the physiological conditions. The present findings suggest that Bcl-2-like proteins play a main role in maintaining of the integrity of cellular components, e.g. mitochondria, rather than in controlling programmed cell death.
Barbieri
D
D
Department of Animal Biology, University of Modena and Reggio Emilia, Modena, Italy.
Malagoli
D
D
Cuoghi
B
B
Ottaviani
E
E
D000818:Animals; D017209:Apoptosis; D002460:Cell Line; D003855:Deoxyribose / Q000494:pharmacology; D018922:HL-60 Cells; D006801:Humans; D007476:Ionophores / Q000494:pharmacology; D019169:Jurkat Cells; D009036:Moths; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism*; D014634:Valinomycin / Q000494:pharmacology
D007476:Ionophores; D019253:Proto-Oncogene Proteins c-bcl-2; D014634:Valinomycin; D003855:Deoxyribose