Laurent F, Benoliel A M, Capo C, Bongrand P
Laboratoire d'Immunologie, Hôpital de Sainte-Marguerite, Marseille, France.
J Leukoc Biol. 1991 Mar;49(3):217-26. doi: 10.1002/jlb.49.3.217.
Different agents such as phorbol myristate acetate (PMA), N-formyl-methionyl-leucylphenylalanine (fMet-Leu-Phe), or opsonized zymosan induced an oxidative burst in rat peritoneal polymorphonuclear leukocytes (PMNs) elicited by casein. Plastic adhesion of PMNs down-regulated superoxide (O2) release stimulated by PMA or fMet-Leu-Phe but had no effect on zymosan-induced O2 generation, indicating that the O2 forming enzyme, the NADPH oxidase, was not affected by modulation and that a common step of the transductional events induced by PMA or fMet-Leu-Phe might be involved in this regulation. We demonstrated that a differential translocation of protein kinase C (PKC) was not responsible for that modulation. PMA-induced secretion of granule content (vitamin B12-binding protein) was not susceptible to modulation, suggesting that the transductional pathways leading to O2 generation and granule secretion are partly separated. The adhesion of PMNs to different substrates (glass, plastic, albumin-, laminin-, fibronectin-, poly-lysine-, or concanavalin A-coated plastic) down-regulated to different extent superoxide release. Whether the nature of the biochemical signal induced by the diverse adhesive stimuli or a physical parameter such as binding strength was involved in this differential behavior remains to be elucidated. Since adhesiveness was dependent on the state of the cytoskeleton and O2 inducers were reported to stimulate actin polymerization, we studied the F-actin content and distribution of PMNs by using the specific fluorescent probe NBD-phallacidin and an original methodology allowing a quantitative analysis of fluorescence on both adherent and suspended cells. PMA induced a polarization of F-actin on suspended PMNs but had no effect on the intracellular distribution of F-actin in adherent PMNs. Thus, we suggest that the adhesion of PMNs induced an immobilization of F-actin, possibly correlated to the down-regulation of one of the transductional pathways involved in the NADPH oxidase activation.
不同的试剂，如佛波酯（PMA）、N-甲酰甲硫氨酰亮氨酰苯丙氨酸（fMet-Leu-Phe）或调理酵母聚糖，可诱导酪蛋白引起的大鼠腹腔多形核白细胞（PMN）产生氧化爆发。PMN的塑料黏附下调了由PMA或fMet-Leu-Phe刺激的超氧化物（O2）释放，但对酵母聚糖诱导的O2生成没有影响，这表明形成O2的酶——NADPH氧化酶不受调节的影响，并且PMA或fMet-Leu-Phe诱导的转导事件的一个共同步骤可能参与了这种调节。我们证明蛋白激酶C（PKC）的差异转位不是这种调节的原因。PMA诱导的颗粒内容物（维生素B12结合蛋白）分泌不易受调节，这表明导致O2生成和颗粒分泌的转导途径部分是分开的。PMN与不同底物（玻璃、塑料、白蛋白、层粘连蛋白、纤连蛋白、聚赖氨酸或伴刀豆球蛋白A包被的塑料）的黏附在不同程度上下调了超氧化物释放。不同黏附刺激诱导的生化信号的性质或诸如结合强度等物理参数是否参与这种差异行为仍有待阐明。由于黏附性取决于细胞骨架的状态，并且据报道O2诱导剂可刺激肌动蛋白聚合，我们使用特异性荧光探针NBD-鬼笔环肽和一种允许对贴壁细胞和悬浮细胞的荧光进行定量分析的原始方法，研究了PMN的F-肌动蛋白含量和分布。PMA诱导悬浮PMN上的F-肌动蛋白极化，但对贴壁PMN中F-肌动蛋白的细胞内分布没有影响。因此，我们认为PMN的黏附诱导了F-肌动蛋白的固定，这可能与NADPH氧化酶激活所涉及的转导途径之一的下调相关。
