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Blood, 1 June 2002, Vol. 99, No. 11, pp. 4039-4047
IMMUNOBIOLOGY
Translocation of the tetraspanin CD63 in association with
human eosinophil mediator release
Salahaddin Mahmudi-Azer,
Gregory P. Downey, and
Redwan Moqbel
From the Pulmonary Research Group, Department of
Medicine, University of Alberta, Edmonton, Canada; and Division of
Respirology, University of Toronto, ON, Canada.
The tetraspanin CD63 (also known as LAMP-3) has been
implicated in phagocytic and intracellular lysosome-phagosome
fusion events. It is also present in eosinophils, with
predominant expression on crystalloid granule membrane. However, its
role in eosinophil function is obscure. We hypothesized that CD63
is associated with intracellular events involved in eosinophil
activation and mediator release. We used a combination of confocal
immunofluorescence microscopy, flow cytometry, and secretion assays,
including  -hexosaminidase, eosinophil peroxidase, and RANTES, to
examine CD63 expression, intracellular localization, and its
association with cell activation and mediator release. In resting
eosinophils, CD63 immunoreactivity was localized to plasma and
crystalloid granule membranes. In interferon- (IFN-)- or
C5a/CB-stimulated cells (10 minutes), intracellular CD63 appeared to
shift to the cell periphery and plasma membrane, while stimulation with
a cocktail of interleukin-3 (IL-3)/IL-5/granulocyte-macrophage
colony-stimulating factor induced the appearance of discrete
intracellular clusters of CD63 immunoreactivity. IFN- induced
mobilization of CD63 to the cell periphery, which coincided with
selective mobilization of RANTES prior to its release, implying CD63
association with piecemeal degranulation. Agonist-induced CD63
mobilization and cell surface up-regulation was associated with
-hexosaminidase, eosinophil peroxidase, and RANTES release. Dexamethasone as well as genistein (a broad-spectrum inhibitor of
tyrosine kinases) inhibited agonist-induced intracellular mobilization of CD63 and RANTES together with cell surface up-regulation of CD63 and
mediator release. This is the first report of an association between
CD63 mobilization and agonist-induced selective mediator release, which
may imply the involvement of CD63 in eosinophil activation and
piecemeal degranulation.
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