The adhesiveness of human polymorphonuclear leukocytes was assessed in serum-coated polystyrene spectrophotometric cuvettes. Capped cuvettes, containing no more than 2 x 10(6) resting or concanavalin A-treated (100 micrograms/ml) polymorphonuclear leukocytes, were laid horizontally and subjected to three 90 degrees rotations on their major axis at fixed times. After incubation at room temperature, non-adherent cells remaining in suspension were counted on the Coulter counter STKS hematological analyzer. After a 16-min incubation (4 min each side of the cuvette) the adhesion of concanavalin A-activated neutrophils ranged from 98% to 100% and the adhesion of resting neutrophils from 30% to 35% (mean 32.4 +/- 2.2%, n = 10). An 8-min incubation (2 min each side) led to approximately 50% adhesion of concanavalin A-activated neutrophils (mean 49.9 +/- 2.2%, range 46%-54%, n = 16), whereas the adhesion of resting cells was about 21% (mean 21.4 +/- 1.6%, range 19%-24%, n = 16). The variation in percentage adhesion in repeated assays did not exceed 4% using concanavalin A-activated cells and 7.5% with resting neutrophils. The procedure is very rapid, easy to perform and precise, and no special apparatus or glassware is necessary. The method also allows microscopic evaluation of shape changes of adherent neutrophils through the clear sides of the cuvettes.
A simple assessment of human neutrophil adhesiveness
BELLAVITE, Paolo;LIPPI, Giuseppe
1994-01-01
Abstract
The adhesiveness of human polymorphonuclear leukocytes was assessed in serum-coated polystyrene spectrophotometric cuvettes. Capped cuvettes, containing no more than 2 x 10(6) resting or concanavalin A-treated (100 micrograms/ml) polymorphonuclear leukocytes, were laid horizontally and subjected to three 90 degrees rotations on their major axis at fixed times. After incubation at room temperature, non-adherent cells remaining in suspension were counted on the Coulter counter STKS hematological analyzer. After a 16-min incubation (4 min each side of the cuvette) the adhesion of concanavalin A-activated neutrophils ranged from 98% to 100% and the adhesion of resting neutrophils from 30% to 35% (mean 32.4 +/- 2.2%, n = 10). An 8-min incubation (2 min each side) led to approximately 50% adhesion of concanavalin A-activated neutrophils (mean 49.9 +/- 2.2%, range 46%-54%, n = 16), whereas the adhesion of resting cells was about 21% (mean 21.4 +/- 1.6%, range 19%-24%, n = 16). The variation in percentage adhesion in repeated assays did not exceed 4% using concanavalin A-activated cells and 7.5% with resting neutrophils. The procedure is very rapid, easy to perform and precise, and no special apparatus or glassware is necessary. The method also allows microscopic evaluation of shape changes of adherent neutrophils through the clear sides of the cuvettes.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.