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Ann Paediatr Rheum. 2012; 1(Supplement 1): 28-28

The Role of MRP8/14 in Cellular Dynamics of Phagocytes

Robiya Joseph, Marc Wolf, Johannes Roth, Thomas Vogl.

Background: The major calcium-binding proteins expressed in phagocytes are myeloid-related protein 8 (MRP8 [S100A8]) and MRP14 (S100A9), two members of the S100 protein family. Phagocytes have the ability to migrate rapidly along the activated endothelium to sites of inflammation. During the process, they adhere onto the endothelium and finally transmigrate through it to inflammatory sites. Since MRP8 and MRP14 are abundantly expressed by these cells, these proteins might possibly be involved in the regulation of these processes.

Methods: Adhesion assay, filter based transmigration assay, flow cytometry, western blotting, RT-PCR.

Results: MRP14 knockout cells spontaneously adhered to a greater extend and transmigrated more in number than wildtype cells. Wildtype cells showed a higher basal surface expression of CD49d (VLA-4α) when compared to the knockout. However, after 1 hour of adhesion, the CD49d expression level was downregulated in the wildtype and upregulated in the knockout cells. Accordingly, adhesion patterns of wildtype and knockout cells reversed in the presence of primary ligands of CD49d e.g. Fibronectin or VCAM-1. Upon blocking CD49d expression, the adhesion and transmigration ability of the MRP8/14 knockout cells decreased. Src and Syk kinases were found to be dysregulated in MRP14 knockout cells compared to wildtype cells. MRP14 knockout cells express more Src but less Syk on both on the total level as well as the activated kinases. These kinases were found to be significantly involved in adhesion and transmigration of both the genotypes as inhibiting the activity of these kinases drastically reduced the efficiency of these two processes.

Conclusion: Our data precisely indicates that MRP8/14 definitely modulates the function of migrating phagocytes and successfully guides their path along the activated endothelium by a mechanism that involves the active participation of the VLA-4α chain wherein the downstream signaling is mediated by the tyrosine kinases Src and Syk.

Key words: MRP8/14, Cellular Dynamics

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