Chemiluminescence and chemotaxis assay of porcine polymorphonuclear cells. A methodological study.
A Boyden-Chamber technique for leukocyte migration was adapted to a polymorphonuclear (PMN) cell concentration of 5 x 10(6) cells/ml an optimal amount of cells for the leading front technique. Casein (0.1%) is so far the most effective chemotactic substance for porcine granulocytes. A formylpeptide++, FMLP (in concentrations of 10(-2) to 10(-10) mmol/l), does not stimulate migration in porcine granulocytes when compared to random migration. For the chemiluminescence assay a concentration of 5 x 10(6) PMN cells/ml buffer is needed to obtain optimal conditions in pig granulocytes. Moreover the chemiluminescence should be performed within a standardized interval after the isolation of leukocytes since the cell activity declines rapidly after the isolation. The chemiluminescence and migration assays were standardized at 37 degrees C; the activity of the granulocytes did not show any significant variation within a temperature interval of 37-39 degrees C, whereas temperatures below 37 degrees C significantly reduce the granulocytic activity.
Thoren-Tolling
K
K
Department of Medicine and Surgery, Veterinary Faculty, University of Agricultural Sciences, Uppsala, Sweden.
D000818:Animals; D002464:Cell Migration Inhibition; D002469:Cell Separation; D002499:Centrifugation, Density Gradient; D002634:Chemotaxis, Leukocyte; D005260:Female; D008163:Luminescent Measurements; D008297:Male; D009504:Neutrophils / Q000276:immunology*; D013552:Swine / Q000276:immunology*