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...

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Bibliographic Details
Published in:PloS one 2016-08, Vol.11 (8), p.e0160685-e0160685
Main Authors: Taylor, Lewis, Brodermann, Maximillian Hugo, McCaffary, David, Iqbal, Asif Jilani, Greaves, David R
Format: Article
Language:English
Subjects:
AKT protein
Animals
Biology and Life Sciences
Blocking antibodies
Bone marrow
Calcium (intracellular)
Calcium - metabolism
Cardiovascular disease
Cell adhesion & migration
Cell culture
Cell Line
Cell migration
Chemokine CCL2 - genetics
Chemokine CCL2 - metabolism
Chemokines
Chemotactic factors
Chemotaxis
Chemotaxis - drug effects
Chemotaxis - physiology
Complement (Immunology)
Complement C5a - metabolism
Complement component C5a
Cytoskeleton
Drug development
Electrical impedance
Exposure
Humans
Immunologic Factors - pharmacology
Inflammation
Inhibition
Intracellular signalling
Laboratories
Macrophages
Macrophages - drug effects
Macrophages - physiology
Male
Medicine and Health Sciences
Mice, Inbred C57BL
Microscopy
Monocyte chemoattractant protein 1
Monocytes
Monocytes - drug effects
Monocytes - physiology
Nerve Growth Factors - chemistry
Nerve Growth Factors - metabolism
Nerve Growth Factors - pharmacology
Netrin-1
Pathology
Penicillin
Phosphatidylinositol 3-Kinases - metabolism
Phosphorylation
Physical Sciences
Real time
Receptor, Anaphylatoxin C5a - metabolism
Receptors
Research and Analysis Methods
Rodents
Signal transduction
Signal Transduction - drug effects
Tumor Suppressor Proteins - chemistry
Tumor Suppressor Proteins - metabolism
Tumor Suppressor Proteins - pharmacology
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Summary: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.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0160685