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CCL4 induces inflammatory signalling and barrier disruption in the neurovascular endothelium

BACKGROUND: During neuroinflammation many chemokines alter the function of the blood-brain barrier (BBB) that regulates the entry of macromolecules and immune cells into the brain. As the milieu of the brain is altered, biochemical and structural changes contribute to the pathogenesis of neuroinflam...

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Autores principales: Estevao, Carolina, Bowers, Chantelle E., Luo, Ding, Sarker, Mosharraf, Hoeh, Alexandra Eva, Frudd, Karen, Turowski, Patric, Greenwood, John
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8560974/
https://www.ncbi.nlm.nih.gov/pubmed/34755124
http://dx.doi.org/10.1016/j.bbih.2021.100370
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author Estevao, Carolina
Bowers, Chantelle E.
Luo, Ding
Sarker, Mosharraf
Hoeh, Alexandra Eva
Frudd, Karen
Turowski, Patric
Greenwood, John
author_facet Estevao, Carolina
Bowers, Chantelle E.
Luo, Ding
Sarker, Mosharraf
Hoeh, Alexandra Eva
Frudd, Karen
Turowski, Patric
Greenwood, John
author_sort Estevao, Carolina
collection PubMed
description BACKGROUND: During neuroinflammation many chemokines alter the function of the blood-brain barrier (BBB) that regulates the entry of macromolecules and immune cells into the brain. As the milieu of the brain is altered, biochemical and structural changes contribute to the pathogenesis of neuroinflammation and may impact on neurogenesis. The chemokine CCL4, previously known as MIP-1β, is upregulated in a wide variety of central nervous system disorders, including multiple sclerosis, where it is thought to play a key role in the neuroinflammatory process. However, the effect of CCL4 on BBB endothelial cells (ECs) is unknown. MATERIALS AND METHODS: Expression and distribution of CCR5, phosphorylated p38, F-actin, zonula occludens-1 (ZO-1) and vascular endothelial cadherin (VE-cadherin) were analysed in the human BBB EC line hCMEC/D3 by Western blot and/or immunofluorescence in the presence and absence of CCL4. Barrier modulation in response to CCL4 using hCMEC/D3 monolayers was assessed by measuring molecular flux of 70 ​kDa RITC-dextran and transendothelial lymphocyte migration. Permeability changes in response to CCL4 in vivo were measured by an occlusion technique in pial microvessels of Wistar rats and by fluorescein angiography in mouse retinae. RESULTS: CCR5, the receptor for CCL4, was expressed in hCMEC/D3 cells. CCL4 stimulation led to phosphorylation of p38 and the formation of actin stress fibres, both indicative of intracellular chemokine signalling. The distribution of junctional proteins was also altered in response to CCL4: junctional ZO-1 was reduced by circa 60% within 60 ​min. In addition, surface VE-cadherin was redistributed through internalisation. Consistent with these changes, CCL4 induced hyperpermeability in vitro and in vivo and increased transmigration of lymphocytes across monolayers of hCMEC/D3 cells. CONCLUSION: These results show that CCL4 can modify BBB function and may contribute to disease pathogenesis.
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spelling pubmed-85609742021-11-08 CCL4 induces inflammatory signalling and barrier disruption in the neurovascular endothelium Estevao, Carolina Bowers, Chantelle E. Luo, Ding Sarker, Mosharraf Hoeh, Alexandra Eva Frudd, Karen Turowski, Patric Greenwood, John Brain Behav Immun Health Full Length Article BACKGROUND: During neuroinflammation many chemokines alter the function of the blood-brain barrier (BBB) that regulates the entry of macromolecules and immune cells into the brain. As the milieu of the brain is altered, biochemical and structural changes contribute to the pathogenesis of neuroinflammation and may impact on neurogenesis. The chemokine CCL4, previously known as MIP-1β, is upregulated in a wide variety of central nervous system disorders, including multiple sclerosis, where it is thought to play a key role in the neuroinflammatory process. However, the effect of CCL4 on BBB endothelial cells (ECs) is unknown. MATERIALS AND METHODS: Expression and distribution of CCR5, phosphorylated p38, F-actin, zonula occludens-1 (ZO-1) and vascular endothelial cadherin (VE-cadherin) were analysed in the human BBB EC line hCMEC/D3 by Western blot and/or immunofluorescence in the presence and absence of CCL4. Barrier modulation in response to CCL4 using hCMEC/D3 monolayers was assessed by measuring molecular flux of 70 ​kDa RITC-dextran and transendothelial lymphocyte migration. Permeability changes in response to CCL4 in vivo were measured by an occlusion technique in pial microvessels of Wistar rats and by fluorescein angiography in mouse retinae. RESULTS: CCR5, the receptor for CCL4, was expressed in hCMEC/D3 cells. CCL4 stimulation led to phosphorylation of p38 and the formation of actin stress fibres, both indicative of intracellular chemokine signalling. The distribution of junctional proteins was also altered in response to CCL4: junctional ZO-1 was reduced by circa 60% within 60 ​min. In addition, surface VE-cadherin was redistributed through internalisation. Consistent with these changes, CCL4 induced hyperpermeability in vitro and in vivo and increased transmigration of lymphocytes across monolayers of hCMEC/D3 cells. CONCLUSION: These results show that CCL4 can modify BBB function and may contribute to disease pathogenesis. Elsevier 2021-10-22 /pmc/articles/PMC8560974/ /pubmed/34755124 http://dx.doi.org/10.1016/j.bbih.2021.100370 Text en © 2021 The Authors. Published by Elsevier Inc. https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Full Length Article
Estevao, Carolina
Bowers, Chantelle E.
Luo, Ding
Sarker, Mosharraf
Hoeh, Alexandra Eva
Frudd, Karen
Turowski, Patric
Greenwood, John
CCL4 induces inflammatory signalling and barrier disruption in the neurovascular endothelium
title CCL4 induces inflammatory signalling and barrier disruption in the neurovascular endothelium
title_full CCL4 induces inflammatory signalling and barrier disruption in the neurovascular endothelium
title_fullStr CCL4 induces inflammatory signalling and barrier disruption in the neurovascular endothelium
title_full_unstemmed CCL4 induces inflammatory signalling and barrier disruption in the neurovascular endothelium
title_short CCL4 induces inflammatory signalling and barrier disruption in the neurovascular endothelium
title_sort ccl4 induces inflammatory signalling and barrier disruption in the neurovascular endothelium
topic Full Length Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8560974/
https://www.ncbi.nlm.nih.gov/pubmed/34755124
http://dx.doi.org/10.1016/j.bbih.2021.100370
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