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1 Department of Cardiovascular Research Center, University of Virginia Health System, Charlottesville, VA, USA; Department of Anesthesiology and Intensive Care Medicine, University of Tuebingen, Tuebingen, Germany
2 Department of Physiology and Biological Physics, University of Virginia Health System, Charlottesville, VA, USA
3 Department of Cardiovascular Research Center, University of Virginia Health System, Charlottesville, VA, USA; Department of Biomedical Engineering, University of Virginia Health System, Charlottesville, VA, USA
4 Department of Cardiovascular Research Center, University of Virginia Health System, Charlottesville, VA, USA; Department of Physiology and Biological Physics, University of Virginia Health System, Charlottesville, VA, USA; Department of Biomedical Engineering, University of Virginia Health System, Charlottesville, VA, USA
* To whom correspondence should be addressed. E-mail: reutershan{at}virginia.edu.
Infiltration of activated neutrophils (polymorphonuclear leukocytes; PMN) into the lung is an important component of the inflammatory response in acute lung injury. The signals required to direct PMN into the different compartments of the lung have not been fully elucidated. In a murine model of LPS-induced lung injury, we investigated the sequential recruitment of PMN into the pulmonary vasculature, lung interstitium, and alveolar space. Mice were exposed to aerosolized LPS and bronchoalveolar lavage fluid (BAL) and lungs were harvested at different time points. We developed a flow cytometry-based technique to assess in-vivo trafficking of PMN in the intravascular and extravascular lung compartments. Aerosolized LPS induced consistent PMN migration into all lung compartments. We found that sequestration in the pulmonary vasculature occurred within the first hour. Transendothelial migration into the interstitial space started one hour after LPS-exposure and increased continuously until a plateau was reached between twelve and 24 hours. Transepithelial migration into the alveolar airspace was delayed, as the first PMN did not appear until two hours after LPS, reaching a peak at 24 hours. Transendothelial migration was partially and transepithelial migration was completely inhibited by pertussis toxin, indicating involvement of G
i-coupled receptors. These findings confirm LPS-induced migration of PMN into the lung. For the first time, distinct transmigration steps into the different lung compartments are characterized in vivo.
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