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Characterisation of TLR4 signalling and function in two brain endothelial cell lines
Author(s) -
Nilsson Olov Karl,
Hellqvist Hedvig
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.711.6
Subject(s) - microbiology and biotechnology , chemokine , tlr4 , tlr2 , p38 mitogen activated protein kinases , cytokine , receptor , blood–brain barrier , signal transduction , phosphorylation , cell culture , biology , chemotaxis , endothelial stem cell , western blot , chemistry , inflammation , mapk/erk pathway , in vitro , immunology , endocrinology , central nervous system , biochemistry , gene , genetics
The blood‐brain barrier (BBB) plays a crucial role in maintaining brain homeostasis. During pathologies such as stroke the function of the BBB is altered into a pro‐inflammatory phenotype and there is increased expression of adhesion molecules, cytokines and chemokines. Toll‐like receptors (TLR), in particular TLR4 and TLR2, are involved in the outcome of experimental stroke manifested by a protective effect in TLR4 and TLR2 knock out animals. In this study we aim to characterize two brain endothelial cell lines, GPNT and bEND5, with regards to TLR4 signalling and function in order for them to be used for in vitro studies of the blood‐brain barrier investigating the role of these receptors in neutrophil migration. The cell lines were exposed to various concentrations of LPS and intracellular signalling molecules were quantified by Western blot and cytokine release by ELISA, respectively. In the rat GPNT cell line a rapid increase (5–15 min) in MAPK p38 phosphorylation was detected. In these cells the release of the chemotactic cytokine CINC‐1 was increased 5‐fold by 1 ng/ml of LPS compared to untreated control. In the bEND5 cells a rapid increase (15 min) in p38 phosphorylation was also detected whereas phosphorylation of JNK could not be observed. These results warrants further investigation of these cell lines for their suitability for in vitro studies of the role of TLR4 signalling in neutrophil transmigration