Rho and Rac, but not ROCK, are required for secretion of human and mouse eosinophil-associated RNases

Clin Exp Allergy. 2019 Feb;49(2):190-198. doi: 10.1111/cea.13292. Epub 2018 Nov 19.

Abstract

Background: Eosinophil-associated RNases (EARs) are stored preformed in eosinophil cytoplasmic secretory granules and have a key role in eosinophil effector functions in host defence and inflammatory disorders. However, the secretion mechanisms of EARs are poorly understood.

Objective: Our study aimed to understand the involvement of cytoskeleton machinery in EAR secretion.

Methods: Fresh human and mouse eosinophils were stimulated with CCL11, and the secretion of enzymatically active EARs was detected using an RNase activity assay. The involvement of cytoskeletal elements or microtubules was probed using specific inhibitors.

Results: We found that dynamic polymerization of microtubules and cytoskeletal elements, such as Rho and Rac, is required for chemokine-mediated EAR secretion from human and mouse eosinophils. However, inhibition of ROCK (Rho-associated protein kinase) increased EAR secretion in human and mouse eosinophils even in the absence of chemokine stimulation, suggesting ROCK negatively regulates EAR secretion.

Conclusions: Collectively, these data suggest a cytoskeleton-dependent mechanism of EAR secretion from eosinophils, findings that are pertinent to host defence, allergy and other eosinophil-associated diseases.

Keywords: ECP; EDN; degranulation; microtubules; signalling.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chemokine CCL11 / genetics
  • Chemokine CCL11 / immunology
  • Eosinophil Cationic Protein / genetics
  • Eosinophil Cationic Protein / immunology*
  • Eosinophils / immunology*
  • Humans
  • Mice
  • Mice, Inbred BALB C
  • Mice, Transgenic
  • rac GTP-Binding Proteins / genetics
  • rac GTP-Binding Proteins / immunology*
  • rho-Associated Kinases / genetics
  • rho-Associated Kinases / immunology*

Substances

  • CCL11 protein, human
  • Ccl11 protein, mouse
  • Chemokine CCL11
  • rho-Associated Kinases
  • Eosinophil Cationic Protein
  • rac GTP-Binding Proteins