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Human peripheral blood eosinophils produce and release interleukin-8 on stimulation with calcium ionophore
R K Braun1, M Franchini, F Erard
1Swiss Institute of Allergy and Asthma Research (SIAF), Davos.
Insights
Human blood eosinophils produce interleukin-8 (IL-8) when stimulated. This finding suggests eosinophils play a role in immune responses and allergic diseases.
Area of Science:
- Immunology
- Cell Biology
Background:
- Eosinophils are key immune cells involved in host defense and allergic inflammation.
- The specific cytokine production profile of eosinophils, particularly regarding interleukin-8 (IL-8), requires further elucidation.
Purpose of the Study:
- To investigate the capacity of human blood eosinophils to produce interleukin-8 (IL-8).
- To explore the regulatory mechanisms and cellular localization of IL-8 production in eosinophils.
Main Methods:
- Purification of human blood eosinophils to high purity (>99%).
- Stimulation of eosinophils with calcium ionophore in vitro.
- Detection of IL-8 protein via enzyme-linked immunosorbent assay (ELISA).
- Analysis of IL-8 messenger RNA (mRNA) using polymerase chain reaction (PCR).
- Immunohistochemistry to determine cytoplasmic IL-8 localization.
Main Results:
- Calcium ionophore stimulation induced significant IL-8 release from purified eosinophils.
- Eosinophil IL-8 production was inhibited by cyclosporin A and cycloheximide, indicating protein synthesis involvement.
- IL-8 mRNA was detected in stimulated eosinophils, confirming transcriptional activity.
- Eosinophils produced less IL-8 than neutrophils but more than T cells upon stimulation.
- Cytoplasmic IL-8 was confirmed within eosinophils using immunohistochemistry.
Conclusions:
- Human blood eosinophils are a source of IL-8 production and release.
- Eosinophil-derived IL-8 may contribute to parasite defense and the pathogenesis of allergic diseases like asthma.
Abstract:
We present evidence that human blood eosinophils produce interleukin (IL)-8 when stimulated with calcium ionophore. Following in vitro culture of 99% pure eosinophils with calcium ionophore, released IL-8 was detectable by enzyme-linked immunosorbent assay in supernatants. Eosinophil IL-8 production was considerably greater than that of IL-3 or granulocyte macrophage colony-stimulating factor. Furthermore, eosinophil production of IL-8 in the presence of calcium ionophore could be inhibited with the immunomodulating agent cyclosporin A and the protein synthesis inhibitor cycloheximide. In addition, following stimulation of highly purified blood eosinophils with calcium ionophore, IL-8 mRNA was detectable after polymerase chain reaction amplification. In comparison with other cells on stimulation with calcium ionophore, eosinophils produce about half as much IL-8 as neutrophils but significantly more than purified T cells. In contrast to monocytes and neutrophils, IL-8 production was not inducible with IL-1 or tumor necrosis factor. Finally, following calcium ionophore stimulation blood eosinophils were shown to contain cytoplasmic IL-8 by employing a monoclonal antibody against IL-8 in conjunction with immunohistochemistry. These observations demonstrate that eosinophils are capable of IL-8 production and release, which may contribute to defense against parasites and to the pathophysiology of allergic and asthmatic disease.
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