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In this study, we investigated the role of extracellular nucleotides in chemokine (KC, MIP-2, MCP-1, and CXCL10) expression and secretion by murine primary intestinal epithelial cells (IECs) with a focus on P2Y 6  receptors. qRT-PCR experiments showed that P2Y 6  was the dominant nucleotide receptor expressed in mouse IEC. In addition, the P2Y 6  ligand UDP induced expression and secretion of CXCL10. For the other studies, we took advantage of mice deficient in P2Y 6  ( P2ry6  -/- ). Similar expression levels of P2Y 1 , P2Y 2 , P2X2, P2X4, and A 2A  were detected in  P2ry6  -/-  and WT IEC. Agonists of TLR3 (poly(I:C)), TLR4 (LPS), P2Y 1 , and P2Y 2  increased the expression and secretion of CXCL10 more prominently in  P2ry6  -/-  IEC than in WT IEC. CXCL10 expression and secretion induced by poly(I:C) in both  P2ry6  -/-  and WT IEC were inhibited by general P2 antagonists (suramin and Reactive-Blue-2), by apyrase, and by specific antagonists of P2Y 1 , P2Y 2 , P2Y 6  (only in WT), and P2X4. Neither adenosine nor an A 2A  antagonist had an effect on CXCL10 expression and secretion. Macrophage chemotaxis was induced by the supernatant of poly(I:C)-treated IEC which was consistent with the level of CXCL10 secreted. Finally, the non-nucleotide agonist FGF2 induced MMP9 mRNA expression also at a higher level in  P2ry6  -/-  IEC than in WT IEC. In conclusion, extracellular nucleotides regulate CXCL10 expression and secretion by IEC. In the absence of P2Y 6 , these effects are modulated by other P2 receptors also present on IEC. These data suggest that the presence of P2Y 6  regulates chemokine secretion and may also regulate IEC homeostasis.

P2Y6 Receptors Regulate CXCL10 Expression and Secretion in Mouse Intestinal Epithelial Cells