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Adhesion and migration of human polymorphonuclear leukocytes across cultured bovine brain microvessel endothelial
K Dorovini-Zis1, P D Bowman, R Prameya
1Department of Pathology, University of British Columbia, Vancouver, Canada.
Abstract:
Adhesion and migration of human polymorphonuclear leukocytes (PMN) across cerebral endothelium were studied in an in vitro model consisting of monolayers of bovine brain microvessel endothelial cells (BBMEC) grown on amniotic stroma or collagen membranes. Polymorphonuclear leukocytes were stimulated to adhere to and migrate across confluent BBMEC monolayers in response to chemotactic gradients produced by formyl-methionyl-leucyl phenylalanine (fMLP), leukotriene B4 (LTB4) or acetyl-glyceryl-ether-phosphorylcholine (AGEPC) placed below the cultures. Under these conditions, PMN adherence to endothelium was 2-10-fold greater than that observed in the absence of chemoattractants or in the presence of equal concentrations of chemoattractants below and above the cultures. Transendothelial migration of PMN occurred rapidly and at focal points across the monolayers. Scanning and electron microscopic studies revealed that stimulated PMN migrated across the monolayers by first adhering to the apical surface of the endothelium and then moving between adjacent endothelial cells. Following their migration, PMN accumulated beneath the endothelium. The overlying endothelial monolayers showed no evidence of disruption and the interendothelial junctions appeared intact at the end of the migration period. We conclude that this in vitro system reproduces the endothelial cell-leukocyte interactions occurring during acute inflammation in vivo and should provide a useful in vitro model for studying the molecular mechanisms underlying these interactions in inflammatory diseases of the central nervous system.
Insights
Human polymorphonuclear leukocytes (PMN) efficiently migrate across brain endothelial cells in vitro. This model aids in understanding leukocyte interactions in central nervous system inflammatory diseases.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Leukocyte adhesion and migration are critical in neuroinflammation.
- Understanding these processes in the central nervous system (CNS) is vital for treating inflammatory diseases.
Purpose of the Study:
- To establish and validate an in vitro model for studying human polymorphonuclear leukocyte (PMN) transmigration across cerebral endothelium.
- To investigate the mechanisms of PMN adhesion and migration in response to specific chemoattractants.
Main Methods:
- Utilized an in vitro model with bovine brain microvessel endothelial cells (BBMEC) monolayers.
- Stimulated PMN adherence and migration using chemotactic gradients of fMLP, LTB4, and AGEPC.
- Employed scanning and transmission electron microscopy to visualize PMN-endothelial interactions.
Main Results:
- Chemoattractants significantly increased PMN adherence (2-10 fold) and transmigration across BBMEC monolayers.
- PMN migrated rapidly through focal points, moving between endothelial cells.
- Endothelial monolayers remained intact, with interendothelial junctions preserved post-migration.
Conclusions:
- The in vitro model accurately replicates in vivo leukocyte-endothelial interactions during acute inflammation in the CNS.
- This system provides a valuable tool for elucidating molecular mechanisms in CNS inflammatory conditions.
- Further research can utilize this model to explore therapeutic targets for neuroinflammatory diseases.