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Novel Inhibition of Monocyte Attachment to Endothelial Cells Involves Hypersulfated Heparan Sulfate
Author(s) -
Ali Mohamed,
Phillips Shane
Publication year - 2012
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.26.1_supplement.680.2
Subject(s) - umbilical vein , heparan sulfate , chemistry , heparanase , monocyte , sulfation , endothelial stem cell , endothelium , protamine sulfate , microbiology and biotechnology , in vitro , biochemistry , cell , protamine , heparin , biology , immunology , endocrinology
The attachment of monocytes to the endothelium involving Heparan Sulfate Proteoglycans (HSPG) is an early hallmark of cardiovascular disease. Previous studies indicate that increased cell surface expression and sulfation of HSPG increases monocyte attachment and contributes to disease. The purpose of this study was to test the hypothesis that protamine derived decapeptides with molecular binding tropism to hyper‐sulfated Heparan Sulfate inhibit monocyte attachment to endothelial cells. We implemented an in vitro endothelial cell flow system with live‐cell time‐lapse microscopy. Human Umbilical Vein Endothelial Cells (HUVECs) were activated with Tumor Necrosis Factor (TNF‐α 15 ng/ml for 24 hours). Monocytic leukemia cells THP‐1 were labeled with Calcein AM for fluorescence microscopy and flow cytometric determination of attachment to HUVEC under laminar flow (5 dynes/cm 2 ). Treatment with marine‐protamines derived cationic peptides (PRTIB‐ONCMY:13‐22) and (PRT1A‐ONCMY:2‐11) at molar concentration of 50μM reduced THP‐1 attachment to HUVECs (60±9% reduction) and preferentially bound to activated HUVECs. Activated HUVECs exhibited increased uptake (35±12%) of the fluorescence labeled peptides compared to controls. We conclude that protamine derived peptides inhibit leukocytes attachment to activated endothelium and may be a novel therapeutic intervention for inflammatory conditions.