Abstract
The blood–brain barrier (BBB) of the central nervous system (CNS) consists of a unique subset of endothelial cells that possess tight junctions which form a relatively impervious physical barrier to a large variety of blood components. Until recently, there have been no good in vitro models for studying the human BBB without the co-culture of feeder cells. The hCMEC/D3 cell line is the first stable, well-differentiated human brain endothelial cell line that grows independently in culture with characteristics that closely resemble those of resident human brain endothelial cells. As our previously published findings demonstrated the importance of adenosine receptor (AR) signaling for lymphocyte entry into the CNS, we wanted to determine if human brain endothelial cells possess the capacity to generate and respond to extracellular adenosine. Utilizing the hCMEC/D3 cell line, we determined that these cells express CD73, the cell surface enzyme that converts extracellular AMP to adenosine. When grown under normal conditions, these cells also express the A1, A2A, and A2B AR subtypes. Additionally, hCMEC/D3 cells are responsive to extracellular AR signaling, as cAMP levels increase following the addition of the broad spectrum AR agonist 5′-N-ethylcarboxamidoadenosine (NECA). Overall, these results indicate that human brain endothelial cells, and most likely the human BBB, have the capacity to synthesize and respond to extracellular adenosine.
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Acknowledgments
We would like to thank the lab of Dr. Eric Denkers and Delbert Abi Abdallah for their expertise in immunoblotting. This work was supported by National Institutes of Health grant: R01 NS063011 (to M.S.B.). Dr. Jeffrey H. Mills holds a Postdoctoral Fellowship from the National Institute of Neurological Disorders and Stroke.
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Mills, J.H., Alabanza, L., Weksler, B.B. et al. Human brain endothelial cells are responsive to adenosine receptor activation. Purinergic Signalling 7, 265–273 (2011). https://doi.org/10.1007/s11302-011-9222-2
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DOI: https://doi.org/10.1007/s11302-011-9222-2