Protease activated receptor signaling is required for African trypanosome traversal of human brain microvascular endothelial cells.
<h4>Background</h4>Using human brain microvascular endothelial cells (HBMECs) as an in vitro model for how African trypanosomes cross the human blood-brain barrier (BBB) we recently reported that the parasites cross the BBB by generating calcium activation signals in HBMECs through the a...
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2009
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oai:doaj.org-article:57eeeeb3c8e24f6f84c9d7feca2de86e2021-11-25T06:33:16ZProtease activated receptor signaling is required for African trypanosome traversal of human brain microvascular endothelial cells.1935-27271935-273510.1371/journal.pntd.0000479https://doaj.org/article/57eeeeb3c8e24f6f84c9d7feca2de86e2009-07-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/19621073/pdf/?tool=EBIhttps://doaj.org/toc/1935-2727https://doaj.org/toc/1935-2735<h4>Background</h4>Using human brain microvascular endothelial cells (HBMECs) as an in vitro model for how African trypanosomes cross the human blood-brain barrier (BBB) we recently reported that the parasites cross the BBB by generating calcium activation signals in HBMECs through the activity of parasite cysteine proteases, particularly cathepsin L (brucipain). In the current study, we examined the possible role of a class of protease stimulated HBMEC G protein coupled receptors (GPCRs) known as protease activated receptors (PARs) that might be implicated in calcium signaling by African trypanosomes.<h4>Methodology/principal findings</h4>Using RNA interference (RNAi) we found that in vitro PAR-2 gene (F2RL1) expression in HBMEC monolayers could be reduced by over 95%. We also found that the ability of Trypanosoma brucei rhodesiense to cross F2RL1-silenced HBMEC monolayers was reduced (39%-49%) and that HBMECs silenced for F2RL1 maintained control levels of barrier function in the presence of the parasite. Consistent with the role of PAR-2, we found that HBMEC barrier function was also maintained after blockade of Galpha(q) with Pasteurella multocida toxin (PMT). PAR-2 signaling has been shown in other systems to have neuroinflammatory and neuroprotective roles and our data implicate a role for proteases (i.e. brucipain) and PAR-2 in African trypanosome/HBMEC interactions. Using gene-profiling methods to interrogate candidate HBMEC pathways specifically triggered by brucipain, several pathways that potentially link some pathophysiologic processes associated with CNS HAT were identified.<h4>Conclusions/significance</h4>Together, the data support a role, in part, for GPCRs as molecular targets for parasite proteases that lead to the activation of Galpha(q)-mediated calcium signaling. The consequence of these events is predicted to be increased permeability of the BBB to parasite transmigration and the initiation of neuroinflammation, events precursory to CNS disease.Dennis J GrabJose C Garcia-GarciaOlga V NikolskaiaYuri V KimAmanda BrownCarlos A PardoYongqing ZhangKevin G BeckerBrenda A WilsonAna Paula C de A LimaJulio ScharfsteinJ Stephen DumlerPublic Library of Science (PLoS)articleArctic medicine. Tropical medicineRC955-962Public aspects of medicineRA1-1270ENPLoS Neglected Tropical Diseases, Vol 3, Iss 7, p e479 (2009) |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Dennis J Grab Jose C Garcia-Garcia Olga V Nikolskaia Yuri V Kim Amanda Brown Carlos A Pardo Yongqing Zhang Kevin G Becker Brenda A Wilson Ana Paula C de A Lima Julio Scharfstein J Stephen Dumler Protease activated receptor signaling is required for African trypanosome traversal of human brain microvascular endothelial cells. |
| description |
<h4>Background</h4>Using human brain microvascular endothelial cells (HBMECs) as an in vitro model for how African trypanosomes cross the human blood-brain barrier (BBB) we recently reported that the parasites cross the BBB by generating calcium activation signals in HBMECs through the activity of parasite cysteine proteases, particularly cathepsin L (brucipain). In the current study, we examined the possible role of a class of protease stimulated HBMEC G protein coupled receptors (GPCRs) known as protease activated receptors (PARs) that might be implicated in calcium signaling by African trypanosomes.<h4>Methodology/principal findings</h4>Using RNA interference (RNAi) we found that in vitro PAR-2 gene (F2RL1) expression in HBMEC monolayers could be reduced by over 95%. We also found that the ability of Trypanosoma brucei rhodesiense to cross F2RL1-silenced HBMEC monolayers was reduced (39%-49%) and that HBMECs silenced for F2RL1 maintained control levels of barrier function in the presence of the parasite. Consistent with the role of PAR-2, we found that HBMEC barrier function was also maintained after blockade of Galpha(q) with Pasteurella multocida toxin (PMT). PAR-2 signaling has been shown in other systems to have neuroinflammatory and neuroprotective roles and our data implicate a role for proteases (i.e. brucipain) and PAR-2 in African trypanosome/HBMEC interactions. Using gene-profiling methods to interrogate candidate HBMEC pathways specifically triggered by brucipain, several pathways that potentially link some pathophysiologic processes associated with CNS HAT were identified.<h4>Conclusions/significance</h4>Together, the data support a role, in part, for GPCRs as molecular targets for parasite proteases that lead to the activation of Galpha(q)-mediated calcium signaling. The consequence of these events is predicted to be increased permeability of the BBB to parasite transmigration and the initiation of neuroinflammation, events precursory to CNS disease. |
| format |
article |
| author |
Dennis J Grab Jose C Garcia-Garcia Olga V Nikolskaia Yuri V Kim Amanda Brown Carlos A Pardo Yongqing Zhang Kevin G Becker Brenda A Wilson Ana Paula C de A Lima Julio Scharfstein J Stephen Dumler |
| author_facet |
Dennis J Grab Jose C Garcia-Garcia Olga V Nikolskaia Yuri V Kim Amanda Brown Carlos A Pardo Yongqing Zhang Kevin G Becker Brenda A Wilson Ana Paula C de A Lima Julio Scharfstein J Stephen Dumler |
| author_sort |
Dennis J Grab |
| title |
Protease activated receptor signaling is required for African trypanosome traversal of human brain microvascular endothelial cells. |
| title_short |
Protease activated receptor signaling is required for African trypanosome traversal of human brain microvascular endothelial cells. |
| title_full |
Protease activated receptor signaling is required for African trypanosome traversal of human brain microvascular endothelial cells. |
| title_fullStr |
Protease activated receptor signaling is required for African trypanosome traversal of human brain microvascular endothelial cells. |
| title_full_unstemmed |
Protease activated receptor signaling is required for African trypanosome traversal of human brain microvascular endothelial cells. |
| title_sort |
protease activated receptor signaling is required for african trypanosome traversal of human brain microvascular endothelial cells. |
| publisher |
Public Library of Science (PLoS) |
| publishDate |
2009 |
| url |
https://doaj.org/article/57eeeeb3c8e24f6f84c9d7feca2de86e |
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