Lucigenin-induced chemiluminescence in human neutrophils in the process of adherence and chemotactic migration measured in a modified Boyden chamber system

Abstract

To answer the question of whether an oxidative burst in neutrophils occurs in the process of adherence and chemotactic migration, we estimated their lucigenin-induced chemiluminescence (CL) responses in a specially devised Boyden chamber that utilized a vial for CL measurement. Neutrophils placed in the upper compartment immediately after isolation induced lucigenin-dependent CL even in the absence of chemotactic factors in the lower chamber, which was completely scavenged by superoxide dismutase at a concentration of 100 micrograms/ml. This response was suggested to be caused by neutrophils in the process of adherence to a filter. When formyl peptide (FMLP) or zymosan-activated serum (ZAS) was placed in the bottom compartment, they were effective to induce CL in the neutrophils already adhered to the membrane filter. These chemotactic factors induced light emission in neutrophils maximally at the concentration of 10(-7) M for FMLP and 5% for ZAS in the reaction mixture without any more increase in CL being observed above these concentrations. This makes a sharp contrast to the findings observed in the so far used unphysiological experimental system, in which neutrophils are abruptly exposed to chemotactic factors likewise in present study. When FMLP or ZAS was directly added to the cells in a upper compartment, CL increased dose-dependently from 10(-9) to 10(-5) M for FMLP, and from 5 to 50% for ZAS.(ABSTRACT TRUNCATED AT 250 WORDS)