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Prepublished online as a Blood First Edition Paper on October 10, 2002; DOI 10.1182/blood-2002-02-0522.
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Blood, 1 March 2003, Vol. 101, No. 5, pp. 1987-1995
PHAGOCYTES
Tumor necrosis factor  induces a caspase-independent death
pathway in human neutrophils
Nikolai A. Maianski,
Dirk Roos, and
Taco W. Kuijpers
From the Sanquin Research at Central Laboratory of the
Netherlands Blood Transfusion Service, Landsteiner Laboratory, and Emma
Children's Hospital, Academic Medical Center, University of Amsterdam,
Amsterdam, The Netherlands; and Medical Academy, Nizhniy
Novgorod, Russia.
Tumor necrosis factor  (TNF-) is a cytokine with
multiple roles in the immune system, including the induction and
potentiation of cellular functions in neutrophils (PMNs). TNF- also
induces apoptotic signals leading to the activation of several
caspases, which are involved in different steps of the process of cell
death. Inhibition of caspases usually increases cell survival. Here, we
found that inhibition of caspases by the general caspase inhibitor zVAD-fmk did not prevent TNF--induced PMN death. After 6 hours of
incubation, TNF- alone caused PMN death with characteristic apoptotic features (typical morphologic changes, DNA laddering, external phosphatidyl serine [PS] exposure in the plasma membrane, Bax clustering and translocation to the mitochondria, and degradation of mitochondria), which coincided with activation of caspase-8 and
caspase-3. However, in the presence of TNF-, PMNs died even when
caspases were completely inhibited. This type of cell death lacked nuclear features of apoptosis (ie, no DNA laddering but aberrant
hyperlobulated nuclei without typical chromatin condensation) and
demonstrated no Bax redistribution, but it did show mitochondria clustering and plasma membrane PS exposure. In contrast, Fas-triggered PMN apoptosis was completely blocked by zVAD-fmk. Experiments with
scavengers of reactive oxygen species (ROS) and with inhibitors of
mitochondrial respiration, with PMN-derived cytoplasts (which lack
mitochondria) and with PMNs from patients with chronic granulomatous disease (which have impaired nicotinamide adenine dinucleotide phosphate [NADPH] oxidase) indicated that
TNF-/zVAD-fmk-induced cell death depends on mitochondria-derived
ROS. Thus, TNF- can induce a "classical," caspase-dependent and
a "nonclassical" caspase-independent cell death.
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