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Blood, Vol. 96 No. 3 (August 1), 2000:
pp. 1030-1038
Caspase-independent commitment phase to apoptosis in activated
blood T lymphocytes: reversibility at low apoptotic insult
Céline Dumont,
Antoine Dürrbach,
Nicolas Bidère,
Matthieu Rouleau,
Guido Kroemer,
Ghislaine Bernard,
François Hirsch,
Bernard Charpentier,
Santos A. Susin, and
Anna Senik
From the Laboratoire d'Immunologie Cellulaire et de
Transplantation, Laboratoire de l'Apoptose, Cancer et Immunité,
Villejuif, France; Unité INSERM 343, Hôpital de
l'Archet, Nice, France.
Little is known about the mechanisms of programmed death triggered
in T lymphocytes by stimuli that can bypass caspase activation. Anti-CD2 monoclonal antibody and staurosporine are such apoptosis inducers because they operate in the presence of broad-spectrum caspase
inhibitors BOC-D.fmk and Z-VAD.fmk. A system was devised, based on the isolation according to density of activated blood T cells
progressively engaged in the apoptotic process. This allowed definition of a sequence of caspase-dependent and
caspase-independent apoptogenic events that are triggered by anti-CD2
and staurosporine. Thus, a commitment phase to apoptosis was defined
that is entirely caspase independent and that is characterized by cell
volume loss, partial chromatin condensation, and release into the
cytosol and the nucleus of mitochondrial "apoptosis-inducing factor
" (AIF). Committed cells were viable, displayed a high mitochondrial
inner transmembrane potential (  m), and lacked large-scale and
oligonucleosomal DNA fragmentation. Mitochondrial release of AIF was
selective because cytochrome c was retained in mitochondria of the very same cells. Mitochondrial release of cytochrome c occurred later, at
the onset of the execution phase of apoptosis, concurrently with
m collapse, poly (ADP-ribose) polymerase cleavage, and DNA
fragmentation. The apoptogenic events of this commitment phase are
reversible if the strength of the stimulus is low and of short duration.
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