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Blood Center of Wisconsin publish paper on Neutrophil Transmigration using Cellix's Platform |
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J Immunol; 2012, 188: Prepublished online 20 January 2012
Proteinase 3 Contributes to Transendothelial Migration of NB1-Positive Neutrophils
Christopher J. Kuckleburg, Sarah B. Tilkens, Sentot Santoso and Peter J. Newman
Abstract
Neutrophil transmigration requires the localization of neutrophils to endothelial cell junctions, in which receptor-ligand interactions and the action of serine protesases promote leukocyte diapedesis. NB1 (CD177) is a neutrophil-expressed surface molecule that has been reported to bind proteinase 3 (PR3), a serine protease released from activated neutrophils. PR3 has demonstrated proteolytic activity on a number of substrates, including extracellular matrix proteins, although its role in neutrophil transmigration is unknown. Recently, NB1 has been shown to be a heterophilic binding partner for the endothelial cell junctional protein, PECAM-1. Disrupting the interaction between NB1 and PECAM-1 significantly inhibits neutrophil transendothelial cell migration on endothelial cell monolayers. Because NB1 interacts with endothelial cell PECAM-1 at cell junctions where transmigration occurs, we considered that NB1-PR3 interactions may play a role in aiding neutrophil diapedesis. Blocking Abs targeting the heterophilic binding domain of PECAM-1 significantly inhibited transmigration of NB1-positive neutrophils through IL-1beta-stimulated endothelial cell monolayers. PR3 expression and activity were significantly increased on NB1-positive neutrophils following transmigration, whereas neutrophils lacking NB1 demonstrated no increase in PR3. Finally, using selective serine protease inhibitors, we determined that PR3 activity facilitated transmigration of NB1-positive neutrophils under both static and flow conditions. These data demonstrate that PR3 contributes in the selective recruitment of the NB1-positive neutrophil population.
Cellix's VenaFlux Platform and Vena8 Endothelial+ biochips were used in this study to investigate neutrophil adhesion and transmigration under flow conditions. It was concluded in the study (see Fig 5 and Fig 6) that PR3 plays and important role in neutrophil transmigration under both static and flow conditions.
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