Table IV

Effect of mAb to Leukocyte Adhesion Molecules on Leukocyte-induced Changes in HUVEC F-Actin Morphology

Stress fiber–positive HUVEC (%)
PMN treatmentMedium (control)CSLEX1 (anti-sLex)60.3 (anti-CD18)M89D3 (anti–PECAM-1)
LPS-treated HUVEC (5 h)64.3 ± 5.8      16.7 ± 2.5*58.4 ± 6.160.9 ± 6.6
Monocyte treatmentMedium (control)CSLEX1 (anti-sLex)60.3 (anti-CD18)M89D3 (anti–PECAM-1)HP2/1 (anti-VLA-4)
LPS-treated HUVEC (24 h)61.3 ± 4.456.2 ± 3.768.3 ± 4.671.2 ± 7.120.1 ± 2.1*
  • Before the assay, HUVEC monolayers were treated for 5 or 24 h with 0.5 μg/ml E. coli LPS. Leukocytes (PMN or monocytes) were treated for 60 min in the presence or absence of mAb CSLEX1, mAb 60.3, mAb M89D3, or mAb HP2/1 (20 μg/ml). Leukocytes were then added to the monolayers (leukocyte/HUVEC ratio = 6–8:1) and the cells were incubated for 10 min at 37°C. The monolayers were then fixed and stained for F-actin. Coverslips were analyzed for HUVEC actin distribution by two observers. An average of 300– 400 cells were examined on each coverslip. Values are means ± SE of three experiments.  

  • * P < 0.05 (paired t test) compared to controls.