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Articles in PresS, published online ahead of print November 30, 2001
Am J Physiol Lung Cell Mol Physiol, 10.1152/ajplung.00261.2001
Submitted on July 16, 2001
Accepted on October 30, 2001
1 Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, USA
* To whom correspondence should be addressed. E-mail: wagnerem{at}jhmi.edu.
Due to 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 post-capillary 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±9.3µm/sec) and adhesion (1.4±0.3 cells/100 ±m) were monitored in post-capillary venules (33.9±1.3µm diameter). At all time points up to 90 min, these parameters were unaltered in control rats (n=7). In contrast, vessels exposed to 1 ±g/ml 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 M 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.
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