Bültmann B D, Haferkamp O, Eggers H J, Gruler H
Blood Cells. 1984;10(1):79-106.
By means of functional, morphological, and biophysical methods the in vitro interaction of Echo virus, type 9, strain A. Barty with human polymorphonuclear leucocytes (PMNs) was investigated and analyzed by statistical methods. Control cells and virus-treated PMNs (15 min, 37 degrees C; PMN: virus (pfu)-ratio ranging from 1:1 to 1:50) were exposed to a chemotactic gradient (N-formylmethionyl-leucylphenylalanine = f-Met-Leu-Phe, 10(-8) M/mm) in a Zigmond chamber. Whereas the track velocity of the moving PMNs was not affected by the virus, the degree of orientation of virus-treated PMNs declined in a way dependent on the viral dose and on the time of PMN:virus interaction, resulting in a shift from chemotactic to chemokinetic response. This virus-induced order-disorder transition of chemotactic response can be described by a logarithmic law in analogy to the Weber-Fechner law. Parallel to the functional disturbances, virus-induced changes of cell shape, which could be confirmed by additional light and electron microscopy techniques, were also detected using statistical analysis of cytological data (median cell size, anisotropy of cell shape) by means of two-dimensional histograms. To investigate f-Met-Leu-Phe- or/and Echo 9 virus-induced PMN-cell membrane changes, the monomer-excimer technique with pyrenedecanoic acid as fluorescent probe was applied, which gives information about structural changes of the cell membrane. Addition of the chemotactic peptide (10(-8) M) to control PMNs resulted in a higher rate of excimer formation obviously due to the formation of new functional (receptor) units (= activated cell membrane). Echo 9 virus exhibited an opposite effect. Quantitative analysis of these results revealed that the f-Met-Leu-Phe-induced cell membrane changes were extinguished by the addition of 2 pfu Echo 9 virus. So far, we have additional indicators of a virus-induced order-disorder transition of chemotactic response of human PMNs on a molecular biological level.
通过功能、形态学和生物物理方法，研究了埃可病毒9型A. Barty株在体外与人多形核白细胞（PMN）的相互作用，并采用统计方法进行了分析。对照细胞和经病毒处理的PMN（37℃，15分钟；PMN与病毒（pfu）的比例为1:1至1:50）在Zigmond小室中暴露于趋化梯度（N-甲酰甲硫氨酰-亮氨酰-苯丙氨酸=f-Met-Leu-Phe，10⁻⁸M/mm）下。虽然移动的PMN的轨迹速度不受病毒影响，但经病毒处理的PMN的定向程度以依赖于病毒剂量和PMN与病毒相互作用时间的方式下降，导致从趋化反应转变为趋化动力学反应。这种病毒诱导的趋化反应的有序-无序转变可以用类似于韦伯-费希纳定律的对数定律来描述。与功能紊乱平行，通过二维直方图对细胞学数据（细胞大小中位数、细胞形状各向异性）进行统计分析，还检测到了病毒诱导的细胞形状变化，这可以通过额外的光学和电子显微镜技术得到证实。为了研究f-Met-Leu-Phe或/和埃可病毒9诱导的PMN细胞膜变化，应用了以芘癸酸为荧光探针的单体-二聚体技术，该技术可提供有关细胞膜结构变化的信息。向对照PMN中添加趋化肽（10⁻⁸M）会导致明显更高的二聚体形成率，这显然是由于形成了新的功能（受体）单元（=活化的细胞膜）。埃可病毒9表现出相反的效果。对这些结果的定量分析表明，添加2个pfu的埃可病毒9可消除f-Met-Leu-Phe诱导的细胞膜变化。到目前为止，我们在分子生物学水平上还有人类PMN趋化反应病毒诱导的有序-无序转变的其他指标。
