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Molecular mechanisms regulating CD13-mediated adhesion.

Immunology (2014-03-19)
Mallika Ghosh, Claire Gerber, M Mamunur Rahman, Kaitlyn M Vernier, Flavia E Pereira, Jaganathan Subramani, Leslie A Caromile, Linda H Shapiro
ABSTRACT

CD13/Aminopeptidase N is a transmembrane metalloproteinase that is expressed in many tissues where it regulates various cellular functions. In inflammation, CD13 is expressed on myeloid cells, is up-regulated on endothelial cells at sites of inflammation and mediates monocyte/endothelial adhesion by homotypic interactions. In animal models the lack of CD13 alters the profiles of infiltrating inflammatory cells at sites of ischaemic injury. Here, we found that CD13 expression is enriched specifically on the pro-inflammatory subset of monocytes, suggesting that CD13 may regulate trafficking and function of specific subsets of immune cells. To further dissect the mechanisms regulating CD13-dependent trafficking we used the murine model of thioglycollate-induced sterile peritonitis. Peritoneal monocytes, macrophages and dendritic cells were significantly decreased in inflammatory exudates from global CD13(KO) animals when compared with wild-type controls. Furthermore, adoptive transfer of wild-type and CD13(KO) primary myeloid cells, or wild-type myeloid cells pre-treated with CD13-blocking antibodies into thioglycollate-challenged wild-type recipients demonstrated fewer CD13(KO) or treated cells in the lavage, suggesting that CD13 expression confers a competitive advantage in trafficking. Similarly, both wild-type and CD13(KO) cells were reduced in infiltrates in CD13(KO) recipients, confirming that both monocytic and endothelial CD13 contribute to trafficking. Finally, murine monocyte cell lines expressing mouse/human chimeric CD13 molecules demonstrated that the C-terminal domain of the protein mediates CD13 adhesion. Therefore, this work verifies that the altered inflammatory trafficking in CD13(KO) mice is the result of aberrant myeloid cell subset trafficking and further defines the molecular mechanisms underlying this regulation.

MATERIALS
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Product Description

Sigma-Aldrich
PKH67 Green Fluorescent Cell Linker Midi Kit for General Cell Membrane Labeling, Distributed for Phanos Technologies
Sigma-Aldrich
PKH26 Red Fluorescent Cell Linker Mini Kit for General Cell Membrane Labeling, Distributed for Phanos Technologies
Sigma-Aldrich
PKH67 Green Fluorescent Cell Linker Mini Kit for General Cell Membrane Labeling, Distributed for Phanos Technologies
Sigma-Aldrich
Leucine Aminopeptidase, microsomal from porcine kidney, Type VI-S, lyophilized powder, ≥12 units/mg protein (biuret)
Sigma-Aldrich
Leucine Aminopeptidase, microsomal from porcine kidney, Type IV-S, ammonium sulfate suspension, 10-40 units/mg protein (Bradford)
Sigma-Aldrich
PKH26 Red Fluorescent Cell Linker Kit for General Cell Membrane Labeling, Distributed for Phanos Technologies
Sigma-Aldrich
PKH26 Red Fluorescent Cell Linker Midi Kit for General Cell Membrane Labeling, Distributed for Phanos Technologies