Abstract
Endothelial cells that functionally express blood brain barrier (BBB) properties are useful surrogates for studying leukocyte-endothelial cell interactions at the BBB. In this study, we compared two different endothelial cellular models: transfected human brain microvascular endothelial cells (THBMECs) and human umbilical vein endothelial cells (HUVECs). With each grow under optimal conditions, confluent THBMEC cultures showed continuous occludin and ZO-1 immunoreactivity, while HUVEC cultures exhibited punctate ZO-1 expression at sites of cell-cell contact only. Confluent THBMEC cultures on 24-well collagen-coated transwell inserts had significantly higher transendothelial electrical resistance (TEER) and lower solute permeability than HUVECs. Confluent THBMECs were more restrictive for mononuclear cell migration than HUVECs. Only THBMECs utilized abluminal CCL5 to facilitate T-lymphocyte migration in vitro although both THBMECs and HUVECs employed CCL3 to facilitate T cell migration. These data establish baseline conditions for using THBMECs to develop in vitro BBB models for studying leukocyte-endothelial interactions during neuroinflammation.
Highlights
Leukocyte recruitment across the blood brain barrier (BBB) into the perivascular space of the central nervous system (CNS) is a key step in the host defense response to pathogens, as well as in neurological disorders such as multiple sclerosis (MS), trauma, and stroke [1,2,3]
The data in this paper showed that Transfected human brain microvascular endothelial cells (THBMECs) differed from human umbilical vein endothelial cells (HUVECs) in their ability to use abluminal CCL5 to mediate T cell migration
There was a progressive increase in monocyte and T lymphocyte migration across both THBMEC and HUVEC cultures, with higher numbers observed at all time points with confluent HUVECs (Figure 3)
Summary
Leukocyte recruitment across the blood brain barrier (BBB) into the perivascular space of the central nervous system (CNS) is a key step in the host defense response to pathogens, as well as in neurological disorders such as multiple sclerosis (MS), trauma, and stroke [1,2,3]. Because endothelial cells from different vascular beds are uniquely adapted to meet the demands of the underlying tissues [12, 13], data from studying HUVECs may not be directly applicable to leukocyte-endothelial interactions at the BBB. In this regard, differences between BMECs and HUVECs have been reported [14,15,16,17]. The data in this paper showed that THBMECs differed from HUVECs in their ability to use abluminal CCL5 to mediate T cell migration These results characterize specific features that distinguish THBMEC from HUVEC cultures, and will help clarify conditions for the development of models to study leukocyte transmigration across BBB in vitro
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