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J. Biochem, 2004, Vol. 135, No. 6 673-681
© 2004 The Japanese Biochemical Society

BIOCHEMISTRY

Sphingosine 1-Phosphate Inhibits Migration of RBL-2H3 Cells via S1P2: Cross-Talk between Platelets and Mast Cells

Eiko Yokoo1, Yutaka Yatomi*,2, Toshiro Takafuta3, Makoto Osada3, Yoshitaka Okamoto§,1 and Yukio Ozaki3

Departments of 1 Otolaryngology and 3 Laboratory Medicine, University of Yamanashi Faculty of Medicine, 1110 Tamaho, Nakakoma, Yamanashi 409-3898; and 2 Department of Laboratory Medicine, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655

To analyze the involvement in allergic reactions of platelets and sphingosine 1-phosphate (Sph-1-P), a lysophospholipid mediator released from activated platelets, the effects of Sph-1-P and a supernatant prepared from activated platelets on mast cell line RBL-2H3 were examined. Sph-1-P strongly inhibited the migration of both non-stimulated and fibronectin-stimulated RBL-2H3 cells, which was reversed by JTE-013, a specific antagonist of G protein-coupled Sph-1-P receptor S1P2; S1P2 was confirmed to be expressed in these cells. A similar anti-motility effect of Sph-1-P was observed in a phagokinetic assay. Consistent with these results, treatment of RBL-2H3 cells with Sph-1-P resulted in a rounded cell morphology, which was blocked by JTE-013. Under the present conditions, Sph-1-P failed to induce intracellular Ca2+ mobilization or histamine degranulation, responses postulated to be elicited by intracellular Sph-1-P. Importantly, the Sph-1-P effect, i.e., the regulation of RBL-2H3 cell motility, was mimicked by the supernatant (both with and without boiling) prepared from activated platelets, and this effect of the supernatant was also blocked by JTE-013. Our results suggest that the motility of mast cells can be regulated by Sph-1-P and also platelets (which release Sph-1-P), via cell surface receptor S1P2 (not through intracellular Sph-1-P actions, postulated previously in the same cells).

* To whom correspondence should be addressed. Tel: +813-5800-8730, Fax: +81-3-5689-0495, E-mail: yatomiy-lab{at}h.u-tokyo.ac.jp

§ Present address: Department of Otolaryngology, Graduate School of Medicine, Chiba University.


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