Eosinophil major basic protein activates human cord blood mast cells primed with fibroblast membranes by integrin-β1

M. Ben-Zimra, I. Bachelet, M. Seaf, G. J. Gleich, F. Levi-Schaffer*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

Background Mast cell (MC) - eosinophil (Eos) activating cross-talk might be critical for the severity and chronicity of allergy. Among soluble mediators, eosinophil major basic protein (MBP), a hallmark of allergy, is particularly important because it was shown to activate specific MC subtypes. We previously demonstrated that MBP activates IgE-desensitized rat MC and human lung and cord blood-derived MC (CBMC) after priming with fibroblast membranal stem cell factor. However, a distinct mechanism for this activation was missing. Therefore, we aimed to investigate it. Methods Major basic protein-1 activation of CBMC primed with fibroblast-derived membranes (FBM) was measured by β-hexosaminidase and tryptase release. Chemical cross-linking followed by micrometric flow cytometry probed direct interactions. Antibodies neutralized integrin-β1 and recognized its active form. Pertussis toxin (Ptx) was used to decrease integrin-β1 active form expression. Hematopoietic cell kinase (Hck) was identified by immunoprecipitation (IP) and silenced by siRNA. Results Major basic protein-1-induced CBMC activation is mediated partly by MBP1-integrin-β1 interaction on the MC surface. FBM prime CBMC via a G protein, as confirmed by Ptx, to shift integrin-β1 to its active form. Following MBP1 binding, integrin-β1 binds Hck that further transduces the activation signal. MC priming with FBM leads to up-regulation in Hck protein level. MC integrin-β1 neutralization inhibits MBP1-induced activation and uptake. Hck silencing results with reduced MBP1-induced activation. Conclusions Fibroblast-derived membranes, integrin-β1, and Hck are involved in MBP1-induced activation of CBMC and therefore represent a distinct mechanism for this activation. This finding might implicate integrin-β1 and Hck as targets for decreasing MC - Eos activating cross-talk in allergy.

Original languageAmerican English
Pages (from-to)1259-1268
Number of pages10
JournalAllergy: European Journal of Allergy and Clinical Immunology
Volume68
Issue number10
DOIs
StatePublished - Oct 2013

Keywords

  • Hck
  • eosinophil major basic protein-1
  • fibroblast membrane fraction
  • integrin-β1
  • mast cells

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