Characterization of mast-cell tryptase-expressing peripheral blood cells as basophils

J Allergy Clin Immunol. 2002 Feb;109(2):287-93. doi: 10.1067/mai.2002.121454.

Abstract

Background: Mast-cell tryptase is a protease with proinflammatory activity, the expression of which by peripheral blood leukocytes (PBLs) has not been fully characterized.

Objective: We examined tryptase expression in human PBLs to further characterize this tryptase-expressing cell population for lineage and disease association.

Methods: PBLs were fixed, permeabilized, stained with antibodies to tryptase and a panel of mast cell- and basophil-specific markers, and analyzed by means of flow cytometry.

Results: Tryptase expression was restricted to a population of cells that stained positive for IgE and negative for the panel of lineage markers (IgE(+), lin(-)). This IgE(+), lin(-) population did not stain for the mast-cell markers Kit or chymase but did stain for the basophil-specific granule proteins recognized by the 2D7 and BB1 mAbs. Per-cell tryptase expression demonstrated a greater than 100-fold range of expression among donors but did not correlate with disease status (asthma or mastocytosis), FEV(1), or serum tryptase concentration. Tryptase was released by purified basophils after anti-IgE activation.

Conclusions: The phenotype of tryptase-expressing PBLs and their lack of increase in patients with mastocytosis demonstrates that these cells are basophils. Per-cell basophil tryptase expression is highly variable between donors, with some donors expressing levels approaching those of mast cells. As such, anti-tryptase antibodies cannot be used to distinguish these 2 cell types from one another by means of flow cytometry. These results demonstrate that tryptase represents an additional mediator through which basophils may contribute to allergic inflammation.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Asthma / immunology*
  • Basophils / enzymology*
  • Basophils / immunology
  • Flow Cytometry
  • Humans
  • Hypersensitivity / immunology*
  • Leukocytes, Mononuclear / classification
  • Mast Cells / enzymology*
  • Mastocytosis / immunology*
  • Phenotype
  • Serine Endopeptidases / metabolism*
  • Staining and Labeling / methods
  • Tryptases

Substances

  • Serine Endopeptidases
  • Tryptases