Quantitative evaluation of neutrophilic chemotaxis in beagles

A quantitative evaluation of neutrophilic chemotaxis was performed, using neutrophils from 12 healthy Beagles. Dextran sedimentation, differential centrifugation, and hypotonic lysis allowed separation of a neutrophil-rich cell suspension. Chemotaxis was studied, using blind-well chambers with 3-micrometers (APD) polycarbonate filters. A frozen (-80 C) bank of pooled normal canine serum was used as the chemotactant, and the chemotactant properties of the serum were destroyed by heat inactivation at 56 C for 30 minutes. Based on membrane and lower blind-well chamber solution neutrophil counts, a method was devised to distinguish between normal neutrophilic chemotaxis, normal neutrophilic chemotaxis with decreased neutrophilic adherence, and defective neutrophilic chemotaxis.