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Blood, Vol. 93 No. 2 (January 15), 1999:
pp. 694-702
The Biologic Role of Interleukin-8: Functional Analysis and
Expression of CXCR1 and CXCR2 on Human Eosinophils
Holger Petering,
Otto Götze,
Daniela Kimmig,
Regina Smolarski,
Alexander Kapp, and
Jörn Elsner
From the Department of Dermatology and Allergology, Hannover Medical
University, Hannover, Germany; and the Department of Immunology,
Georg-August University of Göttingen, Göttingen, Germany.
Chemokines play an important role in attracting granulocytes into
sites of inflammation. Two chemokine subfamilies differ in their
biologic activity for different granulocyte subsets. Whereas CXC
chemokines such as interleukin-8 (IL-8) activate predominantly neutrophils, CC chemokines such as RANTES and eotaxin activate predominantly eosinophils. However, controversial results have been
published in the past regarding the biologic role of IL-8 in eosinophil
activation, particularly in allergic diseases. In this study, we
investigated the functional evidence and expression of both IL-8
receptors, CXCR1 and CXCR2, on highly purified human eosinophils. In
the first set of experiments, a chemotaxis assay was performed showing
that IL-8 did not induce chemotaxis of eosinophils. In addition, and in
contrast to neutrophils and lymphocytes, IL-8 did not induce a rapid
and transient release of cytosolic free Ca2+
([Ca2+]i) in eosinophils, even after
preincubation with TH1- and TH2-like cytokines. To investigate whether
neutrophil contamination might be responsible for the reported IL-8
effects on eosinophils, neutrophils were added to highly purified
eosinophils from the same donor in different concentrations.
Interestingly, as little as 5% of neutrophil contamination was
sufficient to induce an increase of [Ca2+]i
after stimulation with IL-8. Flow cytometry experiments with monoclonal
antibodies against both IL-8 receptors demonstrated no expression of
CXCR1 and CXCR2 on eosinophils before or after cytokine activation.
Reverse transcriptase-polymerase chain reaction experiments showed that eosinophils, in contrast to neutrophils and
lymphocytes, did not express mRNA for CXCR1 and CXCR2. In summary, this
study clearly demonstrates that CXCR1 and CXCR2 are not expressed on
human eosinophils, even after priming with different bioactive
cytokines. Because the CXC chemokine IL-8 did not induce in vitro
effects on human eosinophils, IL-8 may also not contribute in vivo to
the influx of eosinophil granulocytes into sites of allergic
inflammation. Our results suggest that CC chemokines such as eotaxin,
eotaxin-2, and MCP-4 are predominant for the activation of eosinophils.
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