Netrin-1 Reduces Monocyte and Macrophage Chemotaxis towards the Complement Component C5a.
Netrin-1, acting at its cognate receptor UNC5b, has been previously demonstrated to inhibit CC chemokine-induced immune cell migration. In line with this, we found that netrin-1 was able to inhibit CCL2-induced migration of bone marrow derived macrophages (BMDMs). However, whether netrin-1 is capabl...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2016-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4980032?pdf=render |
| _version_ | 1818057819971125248 |
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| author | Lewis Taylor Maximillian Hugo Brodermann David McCaffary Asif Jilani Iqbal David R Greaves |
| author_facet | Lewis Taylor Maximillian Hugo Brodermann David McCaffary Asif Jilani Iqbal David R Greaves |
| author_sort | Lewis Taylor |
| collection | DOAJ |
| description | Netrin-1, acting at its cognate receptor UNC5b, has been previously demonstrated to inhibit CC chemokine-induced immune cell migration. In line with this, we found that netrin-1 was able to inhibit CCL2-induced migration of bone marrow derived macrophages (BMDMs). However, whether netrin-1 is capable of inhibiting chemotaxis to a broader range of chemoattractants remains largely unexplored. As our initial experiments demonstrated that RAW264.7 and BMDMs expressed high levels of C5a receptor 1 (C5aR1) on their surface, we aimed to determine the effect of netrin-1 exposure on monocyte/macrophage cell migration induced by C5a, a complement peptide that plays a major role in multiple inflammatory pathologies. Treatment of RAW264.7 macrophages, BMDMs and human monocytes with netrin-1 inhibited their chemotaxis towards C5a, as measured using two different real-time methods. This inhibitory effect was found to be dependent on netrin-1 receptor signalling, as an UNC5b blocking antibody was able to reverse netrin-1 inhibition of C5a induced BMDM migration. Treatment of BMDMs with netrin-1 had no effect on C5aR1 proximal signalling events, as surface C5aR1 expression, internalisation and intracellular Ca2+ release following C5aR1 ligation remained unaffected after netrin-1 exposure. We next examined receptor distal events that occur following C5aR1 activation, but found that netrin-1 was unable to inhibit C5a induced phosphorylation of ERK1/2, Akt and p38, pathways important for cellular migration. Furthermore, netrin-1 treatment had no effect on BMDM cytoskeletal rearrangement following C5a stimulation as determined by microscopy and real-time electrical impedance sensing. Taken together these data highlight that netrin-1 inhibits monocyte and macrophage cell migration, but that the mechanism behind this effect remains unresolved. Nevertheless, netrin-1 and its cognate receptors warrant further investigation as they may represent a potential avenue for the development of novel anti-inflammatory therapeutics. |
| first_indexed | 2024-12-10T12:50:48Z |
| format | Article |
| id | doaj.art-fba638b7c3c24b4d98306042326175bb |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-10T12:50:48Z |
| publishDate | 2016-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-fba638b7c3c24b4d98306042326175bb2022-12-22T01:48:16ZengPublic Library of Science (PLoS)PLoS ONE1932-62032016-01-01118e016068510.1371/journal.pone.0160685Netrin-1 Reduces Monocyte and Macrophage Chemotaxis towards the Complement Component C5a.Lewis TaylorMaximillian Hugo BrodermannDavid McCaffaryAsif Jilani IqbalDavid R GreavesNetrin-1, acting at its cognate receptor UNC5b, has been previously demonstrated to inhibit CC chemokine-induced immune cell migration. In line with this, we found that netrin-1 was able to inhibit CCL2-induced migration of bone marrow derived macrophages (BMDMs). However, whether netrin-1 is capable of inhibiting chemotaxis to a broader range of chemoattractants remains largely unexplored. As our initial experiments demonstrated that RAW264.7 and BMDMs expressed high levels of C5a receptor 1 (C5aR1) on their surface, we aimed to determine the effect of netrin-1 exposure on monocyte/macrophage cell migration induced by C5a, a complement peptide that plays a major role in multiple inflammatory pathologies. Treatment of RAW264.7 macrophages, BMDMs and human monocytes with netrin-1 inhibited their chemotaxis towards C5a, as measured using two different real-time methods. This inhibitory effect was found to be dependent on netrin-1 receptor signalling, as an UNC5b blocking antibody was able to reverse netrin-1 inhibition of C5a induced BMDM migration. Treatment of BMDMs with netrin-1 had no effect on C5aR1 proximal signalling events, as surface C5aR1 expression, internalisation and intracellular Ca2+ release following C5aR1 ligation remained unaffected after netrin-1 exposure. We next examined receptor distal events that occur following C5aR1 activation, but found that netrin-1 was unable to inhibit C5a induced phosphorylation of ERK1/2, Akt and p38, pathways important for cellular migration. Furthermore, netrin-1 treatment had no effect on BMDM cytoskeletal rearrangement following C5a stimulation as determined by microscopy and real-time electrical impedance sensing. Taken together these data highlight that netrin-1 inhibits monocyte and macrophage cell migration, but that the mechanism behind this effect remains unresolved. Nevertheless, netrin-1 and its cognate receptors warrant further investigation as they may represent a potential avenue for the development of novel anti-inflammatory therapeutics.http://europepmc.org/articles/PMC4980032?pdf=render |
| spellingShingle | Lewis Taylor Maximillian Hugo Brodermann David McCaffary Asif Jilani Iqbal David R Greaves Netrin-1 Reduces Monocyte and Macrophage Chemotaxis towards the Complement Component C5a. PLoS ONE |
| title | Netrin-1 Reduces Monocyte and Macrophage Chemotaxis towards the Complement Component C5a. |
| title_full | Netrin-1 Reduces Monocyte and Macrophage Chemotaxis towards the Complement Component C5a. |
| title_fullStr | Netrin-1 Reduces Monocyte and Macrophage Chemotaxis towards the Complement Component C5a. |
| title_full_unstemmed | Netrin-1 Reduces Monocyte and Macrophage Chemotaxis towards the Complement Component C5a. |
| title_short | Netrin-1 Reduces Monocyte and Macrophage Chemotaxis towards the Complement Component C5a. |
| title_sort | netrin 1 reduces monocyte and macrophage chemotaxis towards the complement component c5a |
| url | http://europepmc.org/articles/PMC4980032?pdf=render |
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