Craddock P R, White J G, Weisdorf D J, Hammerschmidt D E
Inflammation. 1980 Dec;4(4):381-95. doi: 10.1007/BF00916049.
The pulmonary leukostasis and lung dysfunction associated with intravascular complement activation results from C5a-mediated granulocyte (GR) aggregation, a phenomenon which can be reproduced in vitro using standard nephelometric techniques. To produce a more subjective measure of the extent and rate of GR aggregation responses we added a digital integrator to the system. The validity of this approach was substantiated by the close correlation between the aggregating and chemotactic activities of C5a and N-formyl-methionine-leucine-phenylalanine. Use of this technique enabled us to define the dose-response relationship of the aggregation produced by the cationophore A23187 and the inhibitory effect of tetracaine on divalent cation-dependent aggregation responses. The aggregation produced by these three stimuli does not result primarily from simple cross-linking of surface changes because, unlike the passive cell-cell association produced by the cation poly-L-lysine, it is not inhibited by anionic poly-L-glutamic acid. The importance of microtubules as regulators of GR adhesiveness was substantiated by the inhibitory effects of colchicine (but not lumicolchicine) on aggregation in this system. These data suggest that this integration of light transmission increments is a useful adjunct to this basic technique, whether used as a bioassay for chemotactic stimuli or as a model to study the many factors which regulate GR adhesiveness.
与血管内补体激活相关的肺白细胞淤滞和肺功能障碍是由C5a介导的粒细胞（GR）聚集引起的，这一现象可以在体外使用标准比浊技术重现。为了对GR聚集反应的程度和速率进行更主观的测量，我们在系统中添加了一个数字积分器。C5a和N-甲酰甲硫氨酸-亮氨酸-苯丙氨酸的聚集活性与趋化活性之间的密切相关性证实了这种方法的有效性。使用这种技术使我们能够确定离子载体A23187产生的聚集的剂量反应关系以及丁卡因对二价阳离子依赖性聚集反应的抑制作用。这三种刺激产生的聚集并非主要源于表面变化的简单交联，因为与阳离子聚-L-赖氨酸产生的被动细胞-细胞结合不同，它不受阴离子聚-L-谷氨酸的抑制。秋水仙碱（而非光秋水仙碱）对该系统中聚集的抑制作用证实了微管作为GR黏附调节因子的重要性。这些数据表明，这种光透射增量的积分对于这项基本技术是一种有用的辅助手段，无论是用作趋化刺激的生物测定还是用作研究调节GR黏附的许多因素的模型。
