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Blood, Vol. 94 No. 1 (July 1), 1999:
pp. 23-32
Rapid Mobilization of Intracellularly Stored RANTES in Response to
Interferon- in Human Eosinophils
Paige Lacy,
Salahaddin Mahmudi-Azer,
Ben Bablitz,
Stacey C. Hagen,
Juan R. Velazquez,
S.F. Paul Man, and
Redwan Moqbel
From the Pulmonary Research Group, Department of Medicine, University
of Alberta, Edmonton, Alberta, Canada.
The CC chemokine RANTES is synthesized, stored, and upregulated in
response to interferon- (IFN-) in human peripheral blood eosinophils. In this report, we propose that RANTES is rapidly mobilized from eosinophil crystalloid granules during agonist-induced degranulation. We stimulated purified eosinophils (>99%) from atopic
asthmatics with 500 U/mL IFN- to analyze the kinetics of
mobilization and release of RANTES (0 to 240 minutes). We used subcellular fractionation, immunogold analysis, two-color confocal laser scanning microscopy (CLSM), and enzyme-linked immunosorbent assay
(ELISA) to trace the movement of eosinophil-derived RANTES from
intracellular stores to release. RANTES was rapidly mobilized (10 minutes) and released after 120 minutes of stimulation (80 ± 15 pg/mL
per 2 × 106 cells). RANTES appeared to be stored in at
least two intracellular compartments: the matrix of crystalloid
granules, detected by major basic protein and eosinophil peroxidase
activities, and a specialized small secretory vesicle present in light
membrane fractions. The extragranular RANTES was mobilized more rapidly than that of crystalloid granules during IFN- stimulation. This effect was not observed in eosinophils treated with IFN- ,
interleukin-3 (IL-3), IL-5, granulocyte-macrophage colony-stimulating
factor (GM-CSF), or genistein followed by IFN-. Our findings suggest that RANTES may be mobilized and released by piecemeal degranulation upon stimulation, involving transport through a putative pool of small
secretory vesicles.
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