PMID- 26867675 OWN - NLM STAT- MEDLINE DCOM- 20161005 LR - 20181113 IS - 1742-2094 (Electronic) IS - 1742-2094 (Linking) VI - 13 DP - 2016 Feb 11 TI - TGF-beta1 regulates human brain pericyte inflammatory processes involved in neurovasculature function. PG - 37 LID - 10.1186/s12974-016-0503-0 [doi] LID - 37 AB - BACKGROUND: Transforming growth factor beta 1 (TGFbeta1) is strongly induced following brain injury and polarises microglia to an anti-inflammatory phenotype. Augmentation of TGFbeta1 responses may therefore be beneficial in preventing inflammation in neurological disorders including stroke and neurodegenerative diseases. However, several other cell types display immunogenic potential and identifying the effect of TGFbeta1 on these cells is required to more fully understand its effects on brain inflammation. Pericytes are multifunctional cells which ensheath the brain vasculature and have garnered recent attention with respect to their immunomodulatory potential. Here, we sought to investigate the inflammatory phenotype adopted by TGFbeta1-stimulated human brain pericytes. METHODS: Microarray analysis was performed to examine transcriptome-wide changes in TGFbeta1-stimulated pericytes, and results were validated by qRT-PCR and cytometric bead arrays. Flow cytometry, immunocytochemistry and LDH/Alamar Blue(R) viability assays were utilised to examine phagocytic capacity of human brain pericytes, transcription factor modulation and pericyte health. RESULTS: TGFbeta1 treatment of primary human brain pericytes induced the expression of several inflammatory-related genes (NOX4, COX2, IL6 and MMP2) and attenuated others (IL8, CX3CL1, MCP1 and VCAM1). A synergistic induction of IL-6 was seen with IL-1beta/TGFbeta1 treatment whilst TGFbeta1 attenuated the IL-1beta-induced expression of CX3CL1, MCP-1 and sVCAM-1. TGFbeta1 was found to signal through SMAD2/3 transcription factors but did not modify nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) translocation. Furthermore, TGFbeta1 attenuated the phagocytic ability of pericytes, possibly through downregulation of the scavenger receptors CD36, CD47 and CD68. Whilst TGFbeta did decrease pericyte number, this was due to a reduction in proliferation, not apoptotic death or compromised cell viability. CONCLUSIONS: TGFbeta1 attenuated pericyte expression of key chemokines and adhesion molecules involved in CNS leukocyte trafficking and the modulation of microglial function, as well as reduced the phagocytic ability of pericytes. However, TGFbeta1 also enhanced the expression of classical pro-inflammatory cytokines and enzymes which can disrupt BBB functioning, suggesting that pericytes adopt a phenotype which is neither solely pro- nor anti-inflammatory. Whilst the effects of pericyte modulation by TGFbeta1 in vivo are difficult to infer, the reduction in pericyte proliferation together with the elevated IL-6, MMP-2 and NOX4 and reduced phagocytosis suggests a detrimental action of TGFbeta1 on neurovasculature. FAU - Rustenhoven, Justin AU - Rustenhoven J AD - Department of Pharmacology and Clinical Pharmacology, The University of Auckland, Auckland, 1023, New Zealand. AD - Centre for Brain Research, The University of Auckland, Auckland, 1023, New Zealand. FAU - Aalderink, Miranda AU - Aalderink M AD - Department of Pharmacology and Clinical Pharmacology, The University of Auckland, Auckland, 1023, New Zealand. AD - Centre for Brain Research, The University of Auckland, Auckland, 1023, New Zealand. FAU - Scotter, Emma L AU - Scotter EL AD - Department of Pharmacology and Clinical Pharmacology, The University of Auckland, Auckland, 1023, New Zealand. AD - Centre for Brain Research, The University of Auckland, Auckland, 1023, New Zealand. FAU - Oldfield, Robyn L AU - Oldfield RL AD - Lab Plus, Auckland, 1023, New Zealand. FAU - Bergin, Peter S AU - Bergin PS AD - Centre for Brain Research, The University of Auckland, Auckland, 1023, New Zealand. AD - Auckland City Hospital, Auckland, 1023, New Zealand. FAU - Mee, Edward W AU - Mee EW AD - Centre for Brain Research, The University of Auckland, Auckland, 1023, New Zealand. AD - Auckland City Hospital, Auckland, 1023, New Zealand. FAU - Graham, E Scott AU - Graham ES AD - Department of Pharmacology and Clinical Pharmacology, The University of Auckland, Auckland, 1023, New Zealand. AD - Centre for Brain Research, The University of Auckland, Auckland, 1023, New Zealand. FAU - Faull, Richard L M AU - Faull RL AD - Department of Anatomy, The University of Auckland, Auckland, 1023, New Zealand. AD - Centre for Brain Research, The University of Auckland, Auckland, 1023, New Zealand. FAU - Curtis, Maurice A AU - Curtis MA AD - Department of Anatomy, The University of Auckland, Auckland, 1023, New Zealand. AD - Centre for Brain Research, The University of Auckland, Auckland, 1023, New Zealand. FAU - Park, Thomas I-H AU - Park TI AD - Department of Pharmacology and Clinical Pharmacology, The University of Auckland, Auckland, 1023, New Zealand. AD - Department of Anatomy, The University of Auckland, Auckland, 1023, New Zealand. AD - Centre for Brain Research, The University of Auckland, Auckland, 1023, New Zealand. FAU - Dragunow, Mike AU - Dragunow M AD - Department of Pharmacology and Clinical Pharmacology, The University of Auckland, Auckland, 1023, New Zealand. m.dragunow@auckland.ac.nz. AD - Centre for Brain Research, The University of Auckland, Auckland, 1023, New Zealand. m.dragunow@auckland.ac.nz. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20160211 PL - England TA - J Neuroinflammation JT - Journal of neuroinflammation JID - 101222974 RN - 0 (Culture Media, Conditioned) RN - 0 (Cytokines) RN - 0 (Interleukin-1beta) RN - 0 (NF-kappa B) RN - 0 (Receptors, Scavenger) RN - 0 (Smad2 Protein) RN - 0 (Transforming Growth Factor beta1) RN - 0 (Vascular Cell Adhesion Molecule-1) RN - EC 1.14.99.1 (Cyclooxygenase 2) RN - EC 1.14.99.1 (PTGS2 protein, human) RN - EC 1.6.3.- (NADPH Oxidase 4) RN - EC 1.6.3.- (NADPH Oxidases) RN - EC 1.6.3.- (NOX4 protein, human) RN - EC 3.4.24.24 (Matrix Metalloproteinase 2) SB - IM MH - Brain/*cytology MH - Cell Proliferation/drug effects MH - Cell Survival/drug effects MH - Culture Media, Conditioned/pharmacology MH - Cyclooxygenase 2/metabolism MH - Cytokines/*metabolism MH - Gene Expression Regulation/*drug effects MH - Humans MH - Interleukin-1beta/pharmacology MH - Matrix Metalloproteinase 2/metabolism MH - NADPH Oxidase 4 MH - NADPH Oxidases/metabolism MH - NF-kappa B/metabolism MH - Pericytes/*drug effects MH - Phagocytes/*drug effects MH - Receptors, Scavenger/genetics/metabolism MH - Signal Transduction/drug effects MH - Smad2 Protein/metabolism MH - Time Factors MH - Transforming Growth Factor beta1/*pharmacology MH - Vascular Cell Adhesion Molecule-1/metabolism PMC - PMC4751726 EDAT- 2016/02/13 06:00 MHDA- 2016/10/07 06:00 PMCR- 2016/02/11 CRDT- 2016/02/13 06:00 PHST- 2015/12/14 00:00 [received] PHST- 2016/02/03 00:00 [accepted] PHST- 2016/02/13 06:00 [entrez] PHST- 2016/02/13 06:00 [pubmed] PHST- 2016/10/07 06:00 [medline] PHST- 2016/02/11 00:00 [pmc-release] AID - 10.1186/s12974-016-0503-0 [pii] AID - 503 [pii] AID - 10.1186/s12974-016-0503-0 [doi] PST - epublish SO - J Neuroinflammation. 2016 Feb 11;13:37. doi: 10.1186/s12974-016-0503-0.