Chemokinetic accumulation of human neutrophils on immune complex-coated substrata: analysis at a boundary.

Abstract

The Iocomotory 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 partcoated 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 (RFcT) to the under surface, and that the lowered speed of locomotion is due to tethering of neutrophils by substratum-bound IgGFc.

Measurement of individual cell migration parameters for human tissue cells

“…The accuracy of this approach depends on sampling interval, tracking duration, and the migratory characteristics of cell type: sampling rate must be smaller than persistence time so that persistent behavior at short-time scales is not obscured, while the duration the cell is tracked must be several multiples of its persistence time to accurately observe the full turning behavior of the cell. Also, although the presence of immotile cells has been acknowledged in studies of neutrophil (Wilkinson et al, 1984), macrophage (Farrell et al, 1990), and microvessel endothelial cell migration (Stokes et al, 1991), to our knowledge our work is the first to positively and quantitatively discern immotile from motile tissue cells. The strict application of morphological criteria, such as the presence of a single well-defined lamellipod and uropod easily identifiable on motile white blood cells (Zigmond et al, 1981), is qualitative, rather than quantitative, for tissue cells observed under relatively low magnification.…”

The E8 subfragment of laminin promotes locomotion of myoblasts over extracellular matrix.

“…This algorithm was used because with uneven locomotion "speed and persistence" analyses are not reliable (Dunn, 1983;Wilkinson et al, 1984;Dow et al, 1987;Dunn and Brown, 1987). The value of -rt was constant (5 h) for the data tabulated in this study.…”

Optimal estimation of cell movement indices from the statistical analysis of cell tracking data

“…Recently, several investigations have involved statistical characterization of cell tracks obtained from time-lapse observation of cell movement in an isotropic environment (Wilkinson et al, 1984;Takle and Lackie, 1986;Dunn and Brown, 1987; Burton et al, 1987;de Boisfleury-Chevance et al, 1989;Glasgow et al, 1989;Stokes and Lauffenburger, 1991;Saltzman et al, 1991;Buettner and Pittman, 1991;DiMilla et al, 1992). Common features revealed by cell tracks are that cell movement persists in the same direction over short times, but that significant random directional changes occur over longer times.…”