Inhibition of chemotactic motility and trans-endothelial migration of human neutrophils by sphingosine 1-phosphate

• Published in: FEBS letters 1997-12, Vol.420 (2), p.196-200
• Main Authors: Kawa, Shigeyuki, Kimura, Satoshi, Hakomori, Sen-itiro, Igarashi, Yasuyuki
• Format: Article
• Language: English
• Subjects: Cell Adhesion - drug effects; Cell Movement - drug effects; Cells, Cultured; Ceramides - pharmacology; Chemotaxis - drug effects; CPAE, cow pulmonary artery endothelial cell; Enzyme Inhibitors - pharmacology; FCS, fetal calf serum; fMLP, N-formyl-methionyl-leucyl-phenylalanine; Gold Colloid; Humans; HUVEC, human umbilical vein endothelial cell; Inflammation - metabolism; Interleukin-8 - pharmacology; Lysophospholipids; N-Formylmethionine Leucyl-Phenylalanine - pharmacology; Neutrophil migration; Neutrophil motility; Neutrophils; Phagocytosis; PKC, protein kinase C; PMA, phorbol 12-myristate 13-acetate; Protein Kinase C - antagonists & inhibitors; Sph-1-P, sphingosine 1-phosphate; Sphingosine - analogs & derivatives; Sphingosine - pharmacology; Sphingosine 1-phosphate; TMS, N,N,N-trimethylsphingosine; Umbilical Cord
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/S0014-5793(97)01516-0

In previous studies, we reported that sphingosine 1-phosphate (Sph-1-P) inhibits the chemotactic motility of some cancer cell lines such as mouse melanoma cells, as well as human smooth muscle cells, at a very low concentration, as demonstrated by a transwell migration assay method (Proc. Natl. Acad. Sci. USA 89, 9698, 1992; J. Cell Biol. 130, 193, 1995). In this study, we investigated the effect of Sph-1-P on the chemotactic motility and invasiveness of human neutrophils, utilizing three different assay systems: (a) a transwell migration assay where IL-8 or fLMP was added as a chemotactic factor, (b) a phagokinetic assay with gold colloids, and (c) a trans-endothelial migration assay with human umbilical vein endothelial cells (HUVECs) plated on collagen layers. We found that among various sphingosine derivatives, Sph-1-P specifically inhibited the IL-8- or fLMP-induced chemotactic migration of neutrophils at concentrations below 1 μM. Phagokinetic activity of neutrophils was also suppressed by Sph-1-P, but more moderately than by the PKC inhibitory sphingosine analog, trimethylsphingosine. Finally, Sph-1-P inhibited trans-endothelial migration and invasiveness of neutrophils into HUVEC-covered collagen layers, whereas no effect on their adhesion to HUVECs was observed. These observations strongly suggest that Sph-1-P can act as a specific and effective motility regulator of human neutrophils, raising the possibility of future applications of Sph-1-P, or its analogs, as anti-inflammatory agents regulating invasive migration of neutrophils through endothelial layers at injured vascular sites.