Yuli I, Snyderman R
J Clin Invest. 1984 May;73(5):1408-17. doi: 10.1172/JCI111345.
A platelet aggregometer was adapted for the simultaneous measurement of perpendicular light scattering in addition to light transmission. The addition of chemoattractants to polymorphonuclear leukocyte suspensions evoked a single wave of increased light transmission, whereas the perpendicular scattering measurement demonstrated a previously unrecognized biphasic response. The first perpendicular scattering response had no detectable latency and peaked at 10 +/- 1 s, then decayed rapidly. The second response peaked at 40 +/- 5 s, and decayed over several minutes. The dose-response curve of chemoattractants for inducing the rapid (10 +/- 1 s) perpendicular scattering peak corresponded to that which initiated chemotaxis. Initiation of the slow (40 +/- 5 s) peak required 10-fold higher amounts of chemoattractants, and the dose-response curve correlated with the induction of lysosomal enzyme secretion and superoxide anion production. Low doses of aliphatic alcohols, which have been shown to enhance chemotaxis but to inhibit secretion and superoxide anion production, abolished the slow perpendicular light-scattering response but left the fast response intact. Stimulants of secretion induced only slow and prolonged responses that were best observed in transmission measurements. In an attempt to resolve the origin of the light-scattering responses, the morphological changes of polymorphonuclear leukocytes were examined microscopically. Neither aggregation nor morphological whole cell polarization could be correlated with changes in light transmission or perpendicular scattering, which suggested that the source of scattering is of subcellular dimensions. The rapid perpendicular light-scattering response of polymorphonuclear leukocytes to chemoattractants appears to record an initial event in the stimulus-response coupling, and its measurement should provide a useful new tool for the study of leukocyte function. The biphasic nature of the light-scattering responses to chemoattractants, moreover, correlates with the dual regulation of the chemotactic and secretory responses of leukocytes.
血小板凝集仪经过改装,除了能测量光透射外,还能同时测量垂直光散射。向多形核白细胞悬液中添加化学引诱剂会引起光透射增加的单波变化,而垂直散射测量显示出一种先前未被认识到的双相反应。第一个垂直散射反应没有可检测到的延迟,在10±1秒时达到峰值,然后迅速衰减。第二个反应在40±5秒时达到峰值,并在几分钟内衰减。化学引诱剂诱导快速(10±1秒)垂直散射峰值的剂量反应曲线与启动趋化作用的曲线相对应。启动缓慢(40±5秒)峰值需要高10倍的化学引诱剂剂量,且剂量反应曲线与溶酶体酶分泌和超氧阴离子产生的诱导相关。低剂量的脂肪醇已被证明能增强趋化作用,但会抑制分泌和超氧阴离子产生,它消除了缓慢的垂直光散射反应,但使快速反应保持完整。分泌刺激剂仅诱导缓慢且持久的反应,这种反应在透射测量中最易观察到。为了确定光散射反应的起源,对多形核白细胞的形态变化进行了显微镜检查。聚集或形态学上的全细胞极化都与光透射或垂直散射的变化无关,这表明散射源是亚细胞尺寸的。多形核白细胞对化学引诱剂的快速垂直光散射反应似乎记录了刺激-反应偶联中的一个初始事件,其测量应为白细胞功能研究提供一种有用的新工具。此外,对化学引诱剂的光散射反应的双相性质与白细胞趋化和分泌反应的双重调节相关。