Th2 cytokine-primed airway smooth muscle cells induce mast cell chemotaxis via secretion of ATP

Abstract

Objective: Mast cell infiltration into airway smooth muscle (ASM) bundle is an important feature of asthma. Extracellular adenosine triphosphate (eATP) contributes to the initiation of airway inflammation. eATP induces mast cells migration by acting through purinergic receptors. CD39 is an ectonucleotidase that degrades ATP to ADP and AMP. Whether eATP participates in the migration of mast cell towards ASM cells is still unknown. Methods: Airway smooth muscle cells (ASMCs) were isolated from C57/BL6J mice sensitized and challenged with OVA. ASMCs were in vitro cultured and stimulated with IL-4 + IL-13 in the presence or absence of exogenous CD39 or CD39 inhibitor ARL67156. ATP level in the supernatants was measured with ATP content determination kit. CXCL10 concentration in the ASMCs supernatants was measured by ELISA, the mRNA expression of CXCL10 in ASMCs was determined with real-time PCR. Human mast cell line HMC-1 was cultured in Iscove’s-Modified Dubecco’s Medium. The expression of CXCR3 in HMC-1 cells was determined with flow cytometry and real-time PCR, respectively. HMC-1 migration rates were determined with transwell system. Results: In the supernatants of Th2 cytokine-stimulated ASMCs, ATP level was higher than that without stimulation. CD39 decreased, whereas ARL67156 increased ATP level in the supernatants. Both ATP and the supernatants of Th2 cytokine-stimulated ASMCs induced migration of HMC-1 cells. The surface and mRNA expression of CXCR3 in HMC-1 cells, and the mRNA expression and secretion of CXCL10 in ASMCs were increased after stimulation with ATP or Th2 cytokines. All these effects were partially inhibited by CD39. Conclusion: Our data suggested ASMCs in the asthma microenvironment promoted the migration of mast cells via secretion of ATP and the expression of CXCL10/CXCR3 axis. CD39 could reverse this effect and may be a new target for the treatment of asthma.

• Airway smooth muscle cells
• ATP
• CD39
• chemotaxis
• mast cells

Acknowledgements

We thank Dr. J. H. Butterfield (Mayo Clinic, Rochester, MN) for providing us with HMC-1 cell lines.