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Published ahead of print on April 1, 2004, doi:10.1164/rccm.200306-846OC

Am. J. Respir. Crit. Care Med., Volume 170, Number 3, August 2004, 227-233

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Submitted on June 27, 2003
Accepted on March 31, 2004

Pulmonary Neutrophil Infiltration in Murine Sepsis: Role of Inducible Nitric Oxide Synthase

Habib M Razavi1, Le Feng Wang1, Sean Weicker1, Marta Rohan1, Cedrin Law1, David G McCormack1, and Sanjay Mehta1*

1 Departments of Medicine, Physiology and Pharmacology, Division of Respirology and Vascular Biology Group, University of Western Ontario, London Health Sciences Center and Lawson Health Research Institute, London, Ontario, Canada

* To whom correspondence should be addressed. E-mail: david.mccormack{at}lhsc.on.ca.

Nitric oxide (NO) derived from inducible NO synthase (iNOS) contributes to the pathophysiologyof acute lung injury (ALI). The effect of iNOS on pulmonary neutrophil infiltration in ALI is not known. Thus, we assessed pulmonary microvascular neutrophil sequestration, through intravital videomicroscopy, and pulmonary neutrophil infiltration, reflected by myeloperoxidase activity and lavage neutrophil counts, following induction of sepsis by cecal ligation/perforation in wild-type (iNOS+/+) vs. iNOS-/- mice. Pulmonary microvascular neutrophil sequestration was attenuated in septic iNOS-/- vs. iNOS+/+ mice (15±1 vs. 20±1 leukocytes per field, p<0.05), but lavage neutrophil counts were greater in iNOS-/- mice (5.7±1.5 vs. 0.7±0.1%, p<0.05) between six and 18 hours post-CLP. When iNOS+/+ bone-marrow was transplanted into bone-marrow-depleted iNOS-/- mice (+to- chimeras; iNOS limited to marrow-derived inflammatory cells), septic pulmonary microvascular neutrophil sequestration and lavage neutrophil counts were restored to levels seen in septic iNOS+/+ mice. In contrast, in -to+ chimeras, pulmonary neutrophil trafficking was similar to iNOS-/- mice. In vitro cytokine-stimulated neutrophil trans-endothelial migration was significantly greater for iNOS-/- vs. iNOS+/+ neutrophils (7.9±0.7 vs. 3.8±0.6%, p<0.05), but was independent of endothelial iNOS. Thus, neutrophil iNOS-derived NO is an important autocrine modulator of pulmonary neutrophil infiltration in murine sepsis.
Key words: Sepsis, Acute Lung Injury, Neutrophil Infiltration, iNOS, Reciprocal Bone-marrow Transplant Chimeras




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