Reactive oxygen species induce signals that lead to apoptotic DNA degradation in primary CD4+ T cells.
Reactive oxygen species are toxic to cells but they may also have active roles in transducing apoptotic events. To study the role of reactive oxygen species in growth factor depletion induced apoptosis of human primary CD4+ T cells, we used a synthetic manganese porphyrin superoxide dismutase mimetic to detoxify superoxide anions formed during apoptosis. Apoptosis of primary CD4+ T cells was characterized by generation of superoxide anions, plasma membrane phosphatidyl-serine translocation, loss of mitochondrial membrane potential, activation of caspase 3, condensation of chromatin, as well as DNA degradation. The detoxification of superoxide anions did not influence plasma membrane phosphatidyl-serine translocation, or chromatin condensation, and only marginally inhibited the loss of mitochondrial membrane potential and the formation of DNA strand breaks. In contrast, the detoxification of superoxide anions significantly reduced caspase 3 activity and almost completely inhibited the apoptotic decrease in total cellular DNA content as measured by propidium iodide staining. Our results indicate that reactive oxygen anions induce signals leading to efficient DNA degradation after the initial formation of DNA strand breaks. Thus, reactive oxygen anions have active roles in signaling that lead to the apoptotic events.
Pajusto
M
M
Department of Otorhinolaryngology, Helsinki University Central Hospital, Helsinki, Finland.
Toivonen
T H
TH
Tarkkanen
J
J
Jokitalo
E
E
Mattila
P S
PS
D017209:Apoptosis / Q000187:drug effects; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology* / Q000187:drug effects* / Q000201:enzymology / Q000648:ultrastructure; D061945:Caspase Inhibitors; D002469:Cell Separation; D002478:Cells, Cultured; D002675:Child, Preschool; D002843:Chromatin / Q000378:metabolism; D053960:DNA Breaks / Q000187:drug effects; D053938:DNA Fragmentation / Q000187:drug effects*; D004579:Electron Transport / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005434:Flow Cytometry; D006801:Humans; D007223:Infant; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008928:Mitochondria / Q000187:drug effects / Q000201:enzymology / Q000648:ultrastructure; D008933:Mitochondrial Swelling / Q000187:drug effects; D014176:Protein Biosynthesis / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects*; D013481:Superoxides / Q000378:metabolism; D013997:Time Factors
D061945:Caspase Inhibitors; D002843:Chromatin; D004791:Enzyme Inhibitors; D017382:Reactive Oxygen Species; D013481:Superoxides