The biologic role of interleukin-8: functional analysis and expression of CXCR1 and CXCR2 on human eosinophils.
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.
Petering
H
H
Department of Dermatology and Allergology, Hannover Medical University, Hannover, Germany.
Götze
O
O
Kimmig
D
D
Smolarski
R
R
Kapp
A
A
Elsner
J
J
D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000032:analysis* / Q000235:genetics; D002118:Calcium / Q000378:metabolism; D002634:Chemotaxis, Leukocyte; D003600:Cytosol / Q000378:metabolism; D004804:Eosinophils / Q000737:chemistry* / Q000502:physiology; D005434:Flow Cytometry; D006801:Humans; D016209:Interleukin-8 / Q000494:pharmacology / Q000502:physiology*; D009504:Neutrophils; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D019707:Receptors, Chemokine / Q000032:analysis* / Q000235:genetics; D018123:Receptors, Interleukin / Q000032:analysis* / Q000235:genetics; D023062:Receptors, Interleukin-8A; D023063:Receptors, Interleukin-8B
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D016209:Interleukin-8; D012333:RNA, Messenger; D019707:Receptors, Chemokine; D018123:Receptors, Interleukin; D023062:Receptors, Interleukin-8A; D023063:Receptors, Interleukin-8B; D002118:Calcium