The response of human neutrophils to a chemotactic tripeptide (N-formyl-methionyl-leucyl-phenylalanine) studied by microcinematography

Abstract

A quantitative microcinematographic technique for the measurement of cellular motion in response to chemical influences is described. The data are analyzed by using a magnetic digitizer connected to a computer, permitting comparison of several characteristics of such motion. When human granulocytes are placed in a gradient of formyl-methionyl-leucyl-phenylalanine (fMLP) (concentration 10(-6)-10(-8) mol/L) against buffer, the motion of the cells is increased in velocity (chemokinesis) and is directed toward the chemoattractant, as measured by the orientation of the vector of motion. This directionality is confirmed by positive values for the McCutcheon index and analysis of directed linear displacement. Concentrations of fMLP below 10(-9) mol/L did not result in chemokinesis or chemotaxis, presumably due to insufficient stimulus. Concentrations of 10(-6) mol/L fMLP and greater resulted in marked stimulation of the cell surface but reduced directionality and velocity of motion compared with cells in a more optimal gradient. Motion of the unstimulated cells tended to be ortholinear , and this was not increased by the presence of a gradient of fMLP. Hence, as previously shown, the Random Walk analysis, which does not give weight to the direction of motion, is not useful in the analysis of the response of these cells to a chemotaxin in this system.