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Slit-2/Robo-1 modulates the CXCL12/CXCR4-induced chemotaxis of T cells
Anil Prasad1, Zahida Qamri, Jane Wu
1Division of Experimental Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA.
Abstract:
Slit, which mediates its function by binding to the Roundabout (Robo) receptor, has been shown to regulate neuronal, dendritic, and leukocyte migration. However, the molecular mechanism by which the Slit/Robo complex inhibits the migration of cells is not well defined. Here, we showed that Slit-2 can inhibit the CXCL12-induced chemotaxis and transendothelial migration of T cells and monocytes. We observed that CXCR4 associates with Robo-1 and that Slit-2 treatment enhances this association with the Robo-1 receptor. Robo-1 is a single-pass transmembrane receptor whose intracellular region contains four conserved motifs designated as CC0, CC1, CC2, and CC3. Structural and functional analyses of Robo receptors revealed that interaction of the CC3 motif with the CXCR4 receptor may regulate the CXCL12-induced chemotaxis of T cells. We further characterized Slit-2-mediated inhibition of the CXCL12/CXCR4 chemotactic pathway and found that Slit-2 can block the CXCL12-induced activation of the Src and Lck kinases but not Lyn kinase. Although Slit-2 did not inhibit the CXCL12-induced activation of MAPKs, it did inhibit the Akt phosphorylation and Rac activation induced by this chemokine. Altogether, our studies indicate a novel mechanism by which the Slit/Robo complex may inhibit the CXCR4/CXCL12-mediated chemotaxis of T cells.
Insights
Slit-2 protein inhibits T cell and monocyte migration by interacting with the Robo-1 receptor, blocking the CXCL12/CXCR4 signaling pathway and key kinase activations.
Area of Science:
- Immunology
- Cell Biology
- Molecular Biology
Background:
- Slit proteins and Roundabout (Robo) receptors regulate cell migration, including leukocytes.
- The precise molecular mechanisms of Slit/Robo-mediated inhibition of cell migration are not fully understood.
Purpose of the Study:
- To elucidate the molecular mechanism by which Slit-2 inhibits T cell and monocyte chemotaxis and transendothelial migration.
- To investigate the interaction between Slit/Robo complex and the CXCL12/CXCR4 signaling pathway.
Main Methods:
- Investigated Slit-2 effects on T cell and monocyte migration in response to CXCL12.
- Utilized co-immunoprecipitation to assess CXCR4 and Robo-1 receptor association.
- Analyzed the impact of Slit-2 on intracellular signaling pathways, including Src, Lck, MAPKs, Akt, and Rac.
Main Results:
- Slit-2 inhibited CXCL12-induced chemotaxis and transendothelial migration of T cells and monocytes.
- Slit-2 enhanced the association between CXCR4 and Robo-1.
- Slit-2 blocked CXCL12-induced activation of Src and Lck kinases, Akt phosphorylation, and Rac activation, but not MAPK activation.
Conclusions:
- The Slit/Robo complex inhibits T cell chemotaxis via the CXCL12/CXCR4 pathway.
- Interaction between the Robo-1 CC3 motif and CXCR4 may be crucial for regulating T cell chemotaxis.
- Slit-2 acts by disrupting key downstream signaling events in the CXCL12/CXCR4 pathway.
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