Nathan C F
Beatrice and Samuel A. Seaver Laboratory, Department of Medicine, Cornell University Medical College, New York, NY 10021.
J Clin Invest. 1987 Dec;80(6):1550-60. doi: 10.1172/JCI113241.
Recombinant tumor necrosis factor alpha (rTNF alpha) and beta (rTNF beta) did not trigger H2O2 release from PMN in suspension. However, when PMN were plated on polystyrene surfaces coated with serum, fibronectin, vitronectin, laminin, or human umbilical vein endothelial cells (HUVEC), rTNFs induced a massive, prolonged secretory response, similar to that elicited by phorbol myristate acetate (PMA) or bacteria. On serum-coated plates, the maximum sustained rate of H2O2 release in response to rTNF alpha was 2.6 +/- 0.2 nmol/min per 10(6) PMN, the same as that with PMA; release continued for 73 +/- 4 min. On laminin-coated surfaces or HUVEC, release of H2O2 in response to rTNFs was slower, but lasted approximately 3.5 h, reaching the same total (greater than 100 nmol/10(6) PMN). Not only was this response far longer and larger than for other soluble stimuli of the respiratory burst studied with PMN in suspension, but the concentration necessary to elicit a half-maximal response (EC50) for rTNF alpha was orders of magnitude lower (55 pM). Responses were similar with FMLP, but ranged from zero to small with recombinant IFN alpha, recombinant IFN beta, recombinant IFN gamma, platelet-derived growth factor, recombinant IL-1 beta, or bacterial lipopolysaccharide. Adherent monocytes did not secrete H2O2 in response to rTNFs. H2O2 secretion by adherent PMN was first detectable 15-90 min after addition of rTNFs or FMLP. This lag period was unaffected by prior exposure of PMN to rTNF alpha in suspension, by allowing PMN to adhere before adding rTNF alpha, or by incubating adherent PMN in medium conditioned by rTNF alpha-treated PMN. Cytochalasins abolished H2O2 secretion in response to rTNFs, but not FMLP, if added during, but not after, the lag period. Thus, H2O2 secretion from rTNF alpha-treated PMN appears to be a direct but delayed response that requires assembly of microfilaments during exposure to the cytokine. These results suggest that PMN adherent to intra- or extravascular surfaces may undergo a massive, prolonged respiratory burst at the command of macrophages and lymphocytes reacting to microbial products and antigens.
重组肿瘤坏死因子α（rTNFα）和β（rTNFβ）不会引发悬浮状态下中性粒细胞释放过氧化氢（H2O2）。然而，当将中性粒细胞接种在涂有血清、纤连蛋白、玻连蛋白、层粘连蛋白或人脐静脉内皮细胞（HUVEC）的聚苯乙烯表面时，rTNFs会引发大量且持续时间长的分泌反应，类似于佛波酯肉豆蔻酸酯（PMA）或细菌所引发的反应。在涂有血清的平板上，rTNFα刺激下H2O2释放的最大持续速率为每10⁶个中性粒细胞2.6±0.2 nmol/分钟，与PMA刺激时相同；释放持续73±4分钟。在涂有层粘连蛋白的表面或HUVEC上，rTNFs刺激下H2O2的释放较慢，但持续约3.5小时，总量相同（超过100 nmol/10⁶个中性粒细胞）。这种反应不仅比悬浮状态下中性粒细胞对其他呼吸爆发可溶性刺激的反应长得多、大得多，而且rTNFα引发半数最大反应（EC50）所需的浓度要低几个数量级（55 pM）。对甲酰甲硫氨酸-亮氨酸-苯丙氨酸（FMLP）的反应相似，但对重组干扰素α、重组干扰素β、重组干扰素γ、血小板衍生生长因子、重组白细胞介素-1β或细菌脂多糖的反应从零到小不等。贴壁的单核细胞对rTNFs不分泌H2O2。添加rTNFs或FMLP后15 - 90分钟首次检测到贴壁中性粒细胞分泌H2O2。这个延迟期不受中性粒细胞在悬浮状态下预先暴露于rTNFα、在添加rTNFα之前让中性粒细胞贴壁或在rTNFα处理的中性粒细胞条件培养基中培养贴壁中性粒细胞的影响。如果在延迟期内但不是延迟期后添加细胞松弛素，细胞松弛素会消除rTNFs刺激下的H2O2分泌，但不会消除FMLP刺激下的分泌。因此，rTNFα处理的中性粒细胞分泌H2O2似乎是一种直接但延迟的反应，在接触细胞因子期间需要微丝组装。这些结果表明贴附于血管内或血管外表面的中性粒细胞可能在对微生物产物和抗原作出反应的巨噬细胞和淋巴细胞的指令下经历大量且持续时间长的呼吸爆发。
