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Implications for the Regulation of Neutrophil Apoptosis

Andrew J. E. Seely, MD; Daniel E. Swartz, MD; Betty Giannias; Nicolas V. Christou, MD, PhD, FRCSC

Arch Surg. 1998;133:1305-1310.

Objectives  To test the hypothesis that loss of polymorphonuclear neutrophil tumor necrosis factor (TNF-) receptors during transmigration renders the exudate neutrophil refractory to TNF-–mediated stimulation of apoptosis; and to investigate the surface expression of Fas on both circulating and exudate neutrophils.

Design  A prospective cohort study.

Setting  Surgical laboratory of a tertiary care hospital.

Participants  Twenty-one healthy human volunteers.

Interventions  All subjects had circulating neutrophils and exudate neutrophils collected by venipuncture and skin window methods, respectively.

Main Outcome Measures  Circulating and exudate neutrophils were incubated in culture medium (1.0x10⁶neutrophils per milliliter) alone or with TNF- (100 ng/mL). Apoptosis was evaluated by flow cytometry (annexin V–fluorescein isothiocyanate and propidium iodide). Tumor necrosis factor –phycoerythrin and anti–human Fas–fluorescein isothiocyanate were used to evaluate neutrophil TNF- receptors and surface expression of Fas.

Results  Exudate neutrophils had a significant delay in apoptosis rates when compared with circulating neutrophils. The percentage of neutrophils expressing TNF- receptors was significantly diminished after exudation (80%±15% vs 33%±9%; P<.001), as was the median channel number of TNF- phycoerythrin fluorescence (8.1±1.6 vs 5.2±0.5; P=.001). However, the expression of Fas was unchanged after transmigration (percentage positive for Fas: 98.7%±0.7% vs 92.8%±3.4%, P=.89; Fas antibody–fluorescein isothiocyanate median channel fluorescence: 12.2±1.1 vs 13.1±1.2; P=.80). Exposure of exudate neutrophils to TNF- failed to increase their rate of apoptosis.

Conclusions  Exudate polymorphonuclear neutrophils are confirmed to have delayed apoptosis. Loss of TNF- receptors during transmigration is necessary for neutrophil survival in the extravascular inflammatory milieu.

From the Surgical Scientist Program, General Surgery (Drs Seely and Swartz), and Division of General Surgery, Royal Victoria Hospital (Ms Giannias and Dr Christou), McGill University, Montreal, Quebec.

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