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Am J Physiol Heart Circ Physiol 283: H2420-H2430, 2002. First published August 8, 2002; doi:10.1152/ajpheart.00787.2001
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Vol. 283, Issue 6, H2420-H2430, December 2002

Tumor necrosis factor-alpha -induced leukocyte adhesion and microvessel permeability

Min Zeng2, Hong Zhang3, Clifford Lowell3, and Pingnian He1

1 Department of Physiology and Pharmacology, School of Medicine, West Virginia University, Morgantown, West Virginia 26506-9229; 2 Department of Human Physiology, School of Medicine, University of California, Davis 95616; and 3 Department of Laboratory Medicine, University of California, San Francisco, California 94143-0134

The objective of this study was to investigate whether leukocyte adhesion and/or emigration are critical steps in increased microvessel permeability during acute inflammation. To conduct this study, we combined autologous blood perfusion with a single microvessel perfusion technique, which allows microvessel permeability to be measured precisely after the endothelium has interacted with blood-borne stimuli. Experiments were carried out in intact venular microvessels in rat mesenteries. Firm attachment of leukocytes to endothelial cells was induced by intravenous injection of TNF-alpha (3.5 µg/kg) and resuming autoperfusion in a precannulated microvessel. Leukocyte emigration was facilitated by superfusion of formyl-Met-Leu-Phe-OH. Microvessel permeability was measured as hydraulic conductivity (Lp) or the solute permeability coefficient to tetramethylrhodamine isothiocyanate-labeled alpha -lactalbumin before and after leukocyte adhesion and emigration in individually perfused microvessels. We found that perfusion of a microvessel with TNF-alpha did not affect basal microvessel permeability, but intravenous injection of TNF-alpha caused significant leukocyte adhesion. However, the significant leukocyte adhesion and emigration did not cause corresponding increases in either Lp or solute permeability. Thus our results suggest that leukocyte adhesion and emigration do not necessarily increase microvessel permeability and the mechanisms that regulate the adhesion process act independently from mechanisms that regulate permeability. In addition, silver staining of endothelial boundaries demonstrated that leukocytes preferentially adhere at the junctions of endothelial cells. The appearance of the silver lines indicates that the TNF-alpha -induced firm adhesion of leukocyte to microvessel walls did not involve apparent changes in the junctional structure of endothelial cells, which is consistent with the results of permeability measurements.

hydraulic conductivity; permeability coefficient; leukocyte emigration; silver staining; CD11b/CD18 expression


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