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01.12.2018 | Research | Ausgabe 1/2018 Open Access

Journal of Neuroinflammation 1/2018

Unique and shared inflammatory profiles of human brain endothelia and pericytes

Journal of Neuroinflammation > Ausgabe 1/2018
Leon C. D. Smyth, Justin Rustenhoven, Thomas I.-H. Park, Patrick Schweder, Deidre Jansson, Peter A. Heppner, Simon J. O’Carroll, Edward W. Mee, Richard L. M. Faull, Maurice Curtis, Mike Dragunow
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Electronic supplementary material

The online version of this article (https://​doi.​org/​10.​1186/​s12974-018-1167-8) contains supplementary material, which is available to authorized users.



Pericytes and endothelial cells are critical cellular components of the blood-brain barrier (BBB) and play an important role in neuroinflammation. To date, the majority of inflammation-related studies in endothelia and pericytes have been carried out using immortalised cell lines or non-human-derived cells. Whether these are representative of primary human cells is unclear and systematic comparisons of the inflammatory responses of primary human brain-derived pericytes and endothelia has yet to be performed.


To study the effects of neuroinflammation at the BBB, primary brain endothelial cells and pericytes were isolated from human biopsy tissue. Culture purity was examined using qPCR and immunocytochemistry. Electrical cell-substrate impedance sensing (ECIS) was used to determine the barrier properties of endothelial and pericyte cultures. Using immunocytochemistry, cytometric bead array, and ECIS, we compared the responses of endothelia and pericytes to a panel of inflammatory stimuli (IL-1β, TNFα, LPS, IFN-γ, TGF-β1, IL-6, and IL-4). Secretome analysis was performed to identify unique secretions of endothelia and pericytes in response to IL-1β.


Endothelial cells were pure, moderately proliferative, retained the expression of BBB-related junctional proteins and transporters, and generated robust TEER. Both endothelia and pericytes have the same pattern of transcription factor activation in response to inflammatory stimuli but respond differently at the secretion level. Secretome analysis confirmed that endothelia and pericytes have overlapping but distinct secretome profiles in response to IL-1β. We identified several cell-type specific responses, including G-CSF and GM-CSF (endothelial-specific), and IGFBP2 and IGFBP3 (pericyte-specific). Finally, we demonstrated that direct addition of IL-1β, TNFα, LPS, and IL-4 contributed to the loss of endothelial barrier integrity in vitro.


Here, we identify important cell-type differences in the inflammatory response of brain pericytes and endothelia and provide, for the first time, a comprehensive profile of the secretions of primary human brain endothelia and pericytes which has implications for understanding how inflammation affects the cerebrovasculature.
Additional file 2: Full secretome profiler dataset from Fig. 5. Pericytes or endothelia from two different cases were treated with either vehicle, or IL-1β (10 ng/mL) for 24 h before media were harvested and secretions analysed as per Fig. 5. Values represent intensity measurements from each secretion spot (two spots per secretion) from both biological replicates, from all treatment groups (Vehicle/Pericyte, IL-1β/Pericyte, Vehicle/Endothelial, IL-1β/Endothelial). Data are given in an Excel spreadsheet. (XLSX 41 kb)
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