Studies on the regulation of the neutrophil chemotactic response using a rapid and reliable method for measuring random migration and chemotaxis of neutrophil granulocytes

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Abstract
An economic and sensitive test system for measuring random and directional migration of human neutrophils is described. The technique, based on a modified Boyden chamber equipped with a two-filter system, permits a substantial reduction of both incubation time and sample volume. The influence of various technical factors such as the neutrophil concentration in the cell suspension, the incubation time of the chambers, the test concentration of activated plasma or serum, the presence of heparin, and the procedure for separating neutrophils from human peripheral blood, was investigated. Standardized procedures for measuring and reporting neutrophil chemotaxis are proposed. The method has been used to study the significance of factors regulating neutrophil migration such as cytotaxin inactivators and neutrophil immobilizing factors (NIF). Activity of cytotaxin inactivators as assessed in undiluted serum or plasma at pH 7.4, 6.0 or 4.0 was very low. In contrast, potent neutrophil immobilizing activity was found in human serum or diluted plasma. These factors which inhibit migration were accordingly termed neutrophil immobilizing factors of plasma (NIF-P) and neutrophil immobilizing factor of serum (NIF-S). These factors are heat-stable, non-dialysable and of high molecular weight.