Seely A J, Swartz D E, Giannias B, Christou N V
Surgical Scientist Program, General Surgery, Royal Victoria Hospital, Montreal, Quebec, Canada.
Arch Surg. 1998 Dec;133(12):1305-10. doi: 10.1001/archsurg.133.12.1305.
To test the hypothesis that loss of polymorphonuclear neutrophil tumor necrosis factor alpha (TNF-alpha) receptors during transmigration renders the exudate neutrophil refractory to TNF-alpha-mediated stimulation of apoptosis; and to investigate the surface expression of Fas on both circulating and exudate neutrophils.
A prospective cohort study.
Surgical laboratory of a tertiary care hospital.
Twenty-one healthy human volunteers.
All subjects had circulating neutrophils and exudate neutrophils collected by venipuncture and skin window methods, respectively.
Circulating and exudate neutrophils were incubated in culture medium (1.0x10(6) neutrophils per milliliter) alone or with TNF-alpha (100 ng/mL). Apoptosis was evaluated by flow cytometry (annexin V-fluorescein isothiocyanate and propidium iodide). Tumor necrosis factor alpha-phycoerythrin and anti-human Fas-fluorescein isothiocyanate were used to evaluate neutrophil TNF-alpha receptors and surface expression of Fas.
Exudate neutrophils had a significant delay in apoptosis rates when compared with circulating neutrophils. The percentage of neutrophils expressing TNF-alpha receptors was significantly diminished after exudation (80%+/-15% vs 33%+/-9%; P<.001), as was the median channel number of TNF-alpha 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-alpha failed to increase their rate of apoptosis.
Exudate polymorphonuclear neutrophils are confirmed to have delayed apoptosis. Loss of TNF-alpha receptors during transmigration is necessary for neutrophil survival in the extravascular inflammatory milieu.
验证多形核中性粒细胞肿瘤坏死因子α（TNF-α）受体在迁移过程中丢失会使渗出液中的中性粒细胞对TNF-α介导的凋亡刺激产生抗性这一假设；并研究Fas在循环中和渗出液中性粒细胞表面的表达情况。
前瞻性队列研究。
一家三级护理医院的外科实验室。
21名健康人类志愿者。
所有受试者分别通过静脉穿刺和皮肤窗口法采集循环中性粒细胞和渗出液中性粒细胞。
将循环中和渗出液中的中性粒细胞单独或与TNF-α（100 ng/mL）一起在培养基中孵育（每毫升1.0×10⁶个中性粒细胞）。通过流式细胞术（膜联蛋白V-异硫氰酸荧光素和碘化丙啶）评估凋亡情况。使用肿瘤坏死因子α-藻红蛋白和抗人Fas-异硫氰酸荧光素评估中性粒细胞TNF-α受体和Fas的表面表达。
与循环中性粒细胞相比，渗出液中性粒细胞的凋亡率有显著延迟。渗出后，表达TNF-α受体的中性粒细胞百分比显著降低（80%±15%对33%±9%；P<0.001），TNF-α藻红蛋白荧光的中位通道数也显著降低（8.1±1.6对5.2±0.5；P=0.001）。然而，迁移后Fas的表达没有变化（Fas阳性百分比：98.7%±0.7%对92.8%±3.4%，P=0.89；Fas抗体-异硫氰酸荧光素中位通道荧光：12.2±1.1对13.1±1.2；P=0.80）。将渗出液中性粒细胞暴露于TNF-α未能增加其凋亡率。
证实渗出液中的多形核中性粒细胞凋亡延迟。迁移过程中TNF-α受体的丢失是中性粒细胞在血管外炎症环境中存活所必需的。
