Extracellular nucleotides regulate CCL20 release from human primary airway epithelial cells, monocytes and monocyte-derived dendritic cells

• Published in: Journal of cellular physiology 2007-06, Vol.211 (3), p.716-727
• Main Authors: Marcet, Brice, Horckmans, Michael, Libert, Frédérick, Hassid, Sergio, Boeynaems, Jean-Marie, Communi, Didier
• Format: Article
• Language: English
• Subjects: CD4-Positive T-Lymphocytes - cytology; CD4-Positive T-Lymphocytes - metabolism; Cells, Cultured; Chemokine CCL20; Chemokines, CC - genetics; Chemokines, CC - metabolism; Chemotaxis - drug effects; Chemotaxis - immunology; Dendritic Cells - cytology; Dendritic Cells - metabolism; Enzyme Inhibitors - pharmacology; Extracellular Signal-Regulated MAP Kinases - antagonists & inhibitors; Extracellular Signal-Regulated MAP Kinases - metabolism; Extracellular Space - metabolism; Gene Expression - drug effects; Gene Expression - immunology; Humans; Interleukin-8 - metabolism; Lipopolysaccharides - pharmacology; Macrophage Inflammatory Proteins - genetics; Macrophage Inflammatory Proteins - metabolism; Monocytes - cytology; Monocytes - metabolism; Nasal Mucosa - cytology; Nasal Mucosa - immunology; Nasal Mucosa - metabolism; Neutrophils - cytology; Neutrophils - metabolism; NF-kappa B - antagonists & inhibitors; NF-kappa B - metabolism; p38 Mitogen-Activated Protein Kinases - antagonists & inhibitors; p38 Mitogen-Activated Protein Kinases - metabolism; Receptors, Purinergic P2 - genetics; Receptors, Purinergic P2Y2; RNA, Messenger - metabolism; Tumor Necrosis Factor-alpha - pharmacology; Uridine Triphosphate - pharmacology
• ISSN: 0021-9541; 1097-4652
• DOI: 10.1002/jcp.20979

Extracellular nucleotides regulate ion transport and mucociliary clearance in human airway epithelial cells (HAECs) via the activation of P2 receptors, especially P2Y2. Therefore, P2Y2 receptor agonists represent potential pharmacotherapeutic agents to treat cystic fibrosis (CF). Nucleotides also modulate inflammatory properties of immune cells like dendritic cells (DCs), which play an important role in mucosal immunity. Using DNA‐microarray experiments, quantitative RT‐PCR and cytokine measurements, we show here that UTP up‐regulated ∼2‐ to 3‐fold the antimicrobial chemokine CCL20 expression and release in primary HAECs cultured on permeable supports at an air–liquid interface (ALI). Both P2Y2 (ATPγS, UTP, INS365) and P2Y6 (UDP, INS48823) agonists increased CCL20 release. UTP‐induced CCL20 release was insensitive to NF‐κB pathway inhibitors but sensitive to inhibitors of ERK1/2 and p38/MAPK pathways. Furthermore, UTP had no effect on interleukin‐(IL)‐8 release and reduced the release of both CCL20 and IL‐8 induced by TNF‐α and LPS. Accordingly, UTP reduced the capacity of basolateral supernatants of HAECs treated with TNF‐α or LPS to induce the chemoattraction of both CD4+ T lymphocytes and neutrophils. In addition, we show that, in monocyte‐derived DCs, ATPγS, and UDP but not UTP/INS365‐stimulated CCL20 release. Likewise, UDP but not ATPγS was also able to increase CCL20 release from monocytes. Pharmacological experiments suggested an involvement of P2Y11 or P2Y6 receptors through NF‐κB, ERK1/2, and p38/MAPK pathways. Altogether, our data demonstrate that nucleotides may modulate chemokine release and leukocyte recruitment in inflamed airways by acting on both epithelial and immune cells. Our results could be relevant for further clinical investigations in CF. J. Cell. Physiol. 211: 716–727, 2007. © 2007 Wiley‐Liss, Inc.