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Intracellular Localization of Interleukin-6 in Eosinophils From
Atopic Asthmatics and Effects of Interferon 
Paige Lacy,
Francesca Levi-Schaffer,
Salahaddin Mahmudi-Azer,
Ben Bablitz,
Stacey C. Hagen,
Juan Velazquez,
A. Barry Kay, and
Redwan Moqbel
From the Pulmonary Research Group, Department of Medicine, University
of Alberta, Edmonton, Alberta, Canada; and Department of Allergy and
Clinical Immunology, National Heart and Lung Institute, London, UK.
Eosinophils, prominent cells in asthmatic inflammation, have been
shown to synthesize, store, and release an array of up to 18 cytokines
and growth factors, including interleukin-6 (IL-6). In this report, we
show that IL-6 immunofluorescence localizes to the matrix of the
crystalloid granule in peripheral blood eosinophils from atopic
asthmatics using confocal laser scanning microscopy (CLSM). Granule
localization of IL-6 was confirmed using dot-blot analysis and
enzyme-linked immunosorbent assay (ELISA) on subcellular fractions of
highly purified eosinophils produced from density centrifugation across
a 0% to 45% Nycodenz gradient. IL-6 was found to coelute with
eosinophil crystalloid granule marker proteins, including eosinophil
peroxidase (EPO), major basic protein (MBP), arylsulfatase B, and
 -hexosaminidase. Immunoreactivity to IL-6 colocalized with
granule-associated IL-2 and IL-5 in subfractionated eosinophils. We
also made the novel and compelling observation that interferon  (IFN), a Th1-type cytokine, stimulated an early elevation in
eosinophil IL-6 immunoreactivity. A 2.5-fold enhancement of IL-6
immunoreactivity in eosinophil granules was observed within 10 minutes
of IFN treatment (500 U/mL), as determined by subcellular fractionation and CLSM. These findings suggest that IFN has
short-term effects on human eosinophil function and imply that a
physiologic role exists for Th1-type cytokine modulation of Th2-type
responses in these cells.
Blood, Vol. 91 No. 7 (April 1), 1998:
pp. 2508-2516
© 1998 by The American Society of Hematology.
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