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1 Division of Pulmonary and Critical Care Medicine and 2 Division of Clinical Immunology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland 21224
Because of its relative inaccessibility, inflammatory cell extravasation within the airway circulation in vivo has been difficult to investigate in real time. A new method has been established using intravital microscopy in the anesthetized rat to visualize leukocytes in superficial postcapillary venules of the trachea. This technique has been validated using local superfusion of lipopolysaccharide (LPS) and N-formyl-methionyl-leucyl-phenylalanine (FMLP). Basal leukocyte rolling velocity (55.4 ± 9.3 µm/s) and adhesion (1.4 ± 0.3 cells/100 µm) were monitored in postcapillary venules (33.9 ± 1.3 µm diameter). At all time points up to 90 min, these parameters were unaltered in control rats (n = 7). In contrast, vessels exposed to 1 µg/ml of LPS (n = 6) exhibited a 57% reduction in leukocyte rolling velocity and an increase in the number of adherent cells (4.7 ± 1 cells/100 µm, P < 0.05). Superfusion with 0.1 µM of FMLP (n = 6) also resulted in a 45% reduction in rolling velocity and an increase in adherent cells (4 ± 0.7 cells/100 µm, P < 0.05). Histological evaluation confirmed local stimulus-induced leukocyte extravasation. These results demonstrate leukocyte recruitment in the airway microvasculature and provide an important new method to study airway inflammation in real time.
adhesion; inflammation; endotoxin; N-formyl-methionyl-leucyl-phenylalanine
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