Wilkinson P C, Lackie J M, Forrester J V, Dunn G A
J Cell Biol. 1984 Nov;99(5):1761-8. doi: 10.1083/jcb.99.5.1761.
The locomotory behavior of human blood neutrophil leukocytes was studied at a boundary between two surfaces with different chemokinetic properties. This was achieved by time-lapse cinematography of neutrophils moving on coverslips coated with BSA, then part-coated with immune complexes by adding anti-BSA IgG with a straight-line boundary between the BSA and the immune complexes. Cell locomotion was filmed in microscopic fields bisected by the boundary, and kinetic behavior was assessed by comparing speed (orthokinesis), turning behavior (klinokinesis), and the rate of diffusion of the cells on each side of the boundary, using a recently described mathematical analysis of kinesis. In the absence of serum or complement, the proportion of motile cells and their speed and rate of diffusion were greater on BSA than on antiBSA, but there was no consistent difference in turning behavior between cells on the two surfaces. The immune complexes were therefore negatively chemokinetic in comparison with BSA, and this resulted from a negative orthokinesis with little or no contribution from klinokinesis. As would be predicted theoretically, this resulted in gradual accumulation of cells on the immune complexes even in the absence of a chemotactic factor. In further studies, a parallel plate flow chamber was used to show that, under conditions of flow, neutrophils accumulated much more rapidly on a surface coated with BSA-anti-BSA than on BSA alone. Moreover, neutrophils on immune complex-coated surfaces lost their ability to form rosettes with IgG-coated erythrocytes. This suggests that neutrophils on immune complex-coated surfaces redistribute their Fc receptors (RFc gamma) to the under surface, and that the lowered speed of locomotion is due to tethering of neutrophils by substratum-bound IgG-Fc.
在具有不同化学动力学特性的两个表面之间的边界处,研究了人类血液中性粒细胞的运动行为。这是通过对在涂有牛血清白蛋白(BSA)的盖玻片上移动的中性粒细胞进行延时摄影来实现的,然后通过添加抗BSA IgG使部分区域被免疫复合物覆盖,在BSA和免疫复合物之间形成一条直线边界。在被边界一分为二的显微镜视野中拍摄细胞运动,并使用最近描述的运动学数学分析方法,通过比较速度(直线运动)、转向行为(曲折运动)以及边界两侧细胞的扩散速率,来评估动力学行为。在没有血清或补体的情况下,运动细胞的比例及其速度和扩散速率在BSA上比在抗BSA上更大,但两个表面上的细胞在转向行为上没有一致的差异。因此,与BSA相比,免疫复合物具有负化学动力学特性,这是由负直线运动导致的,曲折运动几乎没有或没有贡献。正如理论上所预测的那样,即使在没有趋化因子的情况下,这也导致细胞在免疫复合物上逐渐积累。在进一步的研究中,使用平行板流动腔来表明,在流动条件下,中性粒细胞在涂有BSA - 抗BSA的表面上比在单独的BSA表面上积累得更快。此外,在免疫复合物包被表面上的中性粒细胞失去了与IgG包被的红细胞形成玫瑰花结的能力。这表明在免疫复合物包被表面上的中性粒细胞将其Fc受体(RFcγ)重新分布到下表面,并且运动速度降低是由于中性粒细胞被基质结合的IgG - Fc束缚所致。