Separation and chemiluminescence properties of human, canine and rat polymorphonuclear cells

Human as well as canine and rat polymorphonuclear cells (PMN) were separated from whole blood by centrifugation. Two-step discontinuous Percoll gradients with distinct densities were used. The purity of the preparations was 99.2%, 98.4% and 97.9%, respectively and the corresponding recoveries were 80.1; 66.3% and 69%. The chemiluminescence properties of the isolated PMN and of phagocytes in small quantities of whole blood were compared in luminol-enhanced assays after stimulation with various agents: non-opsonized zymosan (3.5 g/l), phorbol myristate acetate (PMA, 2.8 X 10(-6) M), calcium ionophore A 23187 (10(-5) M) and N-formyl-methionyl-leucyl-phenylalanine (FMLP, 3.5 X 10(-6) M). The isolated cells of the three species responded to all of the various stimuli. Species-related sensitivity followed the order: human greater than canine greater than rat. Responses to the various agents in the human cells was ranked: PMA greater than or equal to A 23187 greater than zymosan greater than FMLP; for the dog: A 23187 greater than PMA greater than zymosan greater than FMLP; and for the rat: zymosan greater than or equal to PMA greater than FMLP greater than or equal to A 23187. Time courses and peak maximum responses were different following stimulation in the absence or presence of autologous plasma. Distinct soluble stimuli resulted in maximum responses below the baseline in the whole blood assays with canine (FMLP) and rat (FMLP, A 23187) phagocytes.