Effect of Anti-ICAM MoAbs on Static Adhesion of Adult Neutrophils to CHO Cell Monolayers
| Neutrophil Treatment . | Treatment of Monolayer . | |||||
|---|---|---|---|---|---|---|
| PBS . | W6/32 . | R6.5Ig . | R6.5fab . | CL18/1 . | CL18/6 . | |
| Unstimulated neutrophils | 7.4 ± 2.7 | 4.7 ± 1.8 | 5.9 ± 2.6 | 4.6 ± 3.2 | 10.4 ± 5.6 | 9.9 ± 4.6 |
| 10 nmol/L fMLP + neutrophils | 37.3 ± 4.3 | 35.8 ± 4.7 | 18.5 ± 1.9* | 20.5 ± 0.8* | 21.1 ± 2.3* | 18.7 ± 2.2* |
| 10 mmol/L fMLP + neutrophils | ||||||
| + PBS | 26.7 ± 3.9 | 23.4 ± 1.6 | 15.1 ± 2.4† | |||
| + W6/32 or R6.5 Fab | 19.2 ± 3.3 | 8.9 ± 1.0‡ | ||||
| Neutrophil Treatment . | Treatment of Monolayer . | |||||
|---|---|---|---|---|---|---|
| PBS . | W6/32 . | R6.5Ig . | R6.5fab . | CL18/1 . | CL18/6 . | |
| Unstimulated neutrophils | 7.4 ± 2.7 | 4.7 ± 1.8 | 5.9 ± 2.6 | 4.6 ± 3.2 | 10.4 ± 5.6 | 9.9 ± 4.6 |
| 10 nmol/L fMLP + neutrophils | 37.3 ± 4.3 | 35.8 ± 4.7 | 18.5 ± 1.9* | 20.5 ± 0.8* | 21.1 ± 2.3* | 18.7 ± 2.2* |
| 10 mmol/L fMLP + neutrophils | ||||||
| + PBS | 26.7 ± 3.9 | 23.4 ± 1.6 | 15.1 ± 2.4† | |||
| + W6/32 or R6.5 Fab | 19.2 ± 3.3 | 8.9 ± 1.0‡ | ||||
Adult neutrophils were allowed to settle for 500 seconds onto monolayers of CHO cells that had been treated with PBS or 10 to 20 μg/mL of control MoAb, W6/32, or anti-ICAM MoAbs for 15 minutes before rinsing and insertion into the Sykes Moore Chamber. All MoAbs were Ig, except for R6.5, in which Fab fragments were also used. The number of interacting neutrophils was counted and averaged per field. The chambers were then inverted for an additional 500 seconds, and the number of neutrophils remaining adherent to the monolayer after this time were counted. Results are expressed as the percentage of settled neutrophils remaining attached after inversion ± SEM (n = 5 to 8). Neutrophils were either unstimulated or stimulated with 10 nmol/L fMLP. In the last set of experiments, no additional MoAb was included with the fMLP and neutrophils or alternatively additional MoAb W6/32 or R6.5 Fab was included in the chambers in which the monolayer had been treated with W6/32 or R6.5 Fab, respectively.
P < .05 v PBS or W6/32 treatment of monolayers.
P < .05 v PBS.
P < .05 v W6/32 treatment of monolayers and neutrophils and R6.5 treatment of monolayers only.