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Pretreatment of Human Polymorphonuclear Leukocytes (PMN) with a New Carbon Monoxide (CO)‐Releasing Molecule (CORM401) Inhibits PMN Migration across Vascular Endothelial Cells
Author(s) -
Inoue Ken,
Woo Martin,
Patterson Eric,
Potter Richard F.,
Capretta Alfredo,
Fraser Douglas D.,
Cepinskas Gediminas
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.634.1
Subject(s) - umbilical vein , chemistry , flow cytometry , confocal microscopy , stimulation , inflammation , granulocyte , in vitro , confocal , lipopolysaccharide , endothelium , biophysics , microbiology and biotechnology , immunology , biochemistry , medicine , biology , geometry , mathematics
Background We have demonstrated that carbon monoxide (CO)‐releasing molecules (CORMs) reduce septic inflammation by interfering with PMN recruitment to the inflamed organs. In this study, we assessed the effects/mechanisms of a newly synthesized Mn 2+ ‐based water‐soluble CORM (CORM401) on PMN adhesion(A)/transendothelial migration(TM) in an experimental model of endotoxemia in vitro. Methods Human umbilical vein endothelial cells (HUVEC) grown on parallel flow channels were stimulated with LPS (1μg/ml; 6 hrs) and interacted with freshly isolated PMN (pretreated for 30 min with CORM401 or its inactive compound, iCORM401; 100μM) for 5 min in the presence of 1.0 dyn/cm 2 shear stress. Subsequently, HUVEC:PMN co‐cultures were additionally perfused for 20 min with medium containing CORM/iCORM401. Video and confocal microscopy were used to analyze PMN A/TM. Results Stimulation of HUVEC with LPS significantly increased PMN A (0.6+/‐0.1 vs 30.9+/‐7.6 cells/mm 2 ; p<0.05) and TM (0.0 vs 41.7+/‐6.8%; p<0.001). Pretreatment of PMN with CORM401 had no effect on PMN A, however significantly reduced PMN TM (41.7+/‐6.8% vs 15.2+/‐4.3; p<0.01). The above findings were confirmed employing confocal microscopy. Conclusion CORM401 interferes with PMN recruitment by specific suppression of PMN migratory potential. (HSFO 393, IRF‐25‐12).