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Blood, Vol. 92 No. 12 (December 15), 1998:
pp. 4808-4818
Involvement of Caspases in Neutrophil Apoptosis: Regulation by
Reactive Oxygen Species
Bengt Fadeel,
Anders Åhlin,
Jan-Inge Henter,
Sten Orrenius, and
Mark B. Hampton
From the Institute of Environmental Medicine, Division of Toxicology,
Karolinska Institutet, Stockholm, Sweden; the Childhood Cancer Research
Unit, Karolinska Hospital, Stockholm, Sweden; the Department of
Pediatrics, Center for Inflammation Research, Karolinska Institutet at
Sach's Children's Hospital and Stockholm Söder Hospital,
Stockholm, Sweden.
Human neutrophils have a short half-life and are believed to die by
apoptosis or programmed cell death both in vivo and in vitro. We found
that caspases are activated in a time-dependent manner in neutrophils
undergoing spontaneous apoptosis, concomitant with other characteristic
features of apoptotic cell death such as morphologic changes,
phosphatidylserine (PS) exposure, and DNA fragmentation. The treatment
of neutrophils with agonistic anti-Fas monoclonal antibodies (MoAbs)
significantly accelerated this process. However, in cells treated with
the potent neutrophil activator phorbol 12-myristate 13-acetate (PMA),
caspase activity was only evident after pharmacologic inhibition of the
nicotinamide adenine dinucleotide phosphate (NADPH)
oxidase. Similarily, inhibition of the NADPH oxidase in constitutive
and Fas/APO-1-triggered apoptosis resulted in increased rather than
suppressed levels of caspase activity, suggesting that reactive oxygen
species may prevent caspases from functioning optimally in these cells.
Moreover, oxidants generated via the NADPH oxidase were essential for
PS exposure during PMA-induced cell death, but not for neutrophils undergoing spontaneous apoptosis. We conclude that caspases are an
important component of constitutive and Fas/APO-1-triggered neutrophil
apoptosis. However, these redox sensitive enzymes are suppressed in
activated neutrophils, and an alternate oxidant-dependent pathway is
used to mediate PS exposure and neutrophil clearance under these
conditions.
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