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Published on: June 23, 2013
The production of macrophage inflammatory protein-1 alpha by human basophils
1Department of Internal Medicine, Division of Allergy and Immunology, University of Texas Medical Branch, Galveston, TX 77555, USA.
Abstract:
Macrophage inflammatory protein-1alpha (MIP-1alpha) has previously been shown to be produced by mononuclear cells, eosinophils, and neutrophils. Its production by basophils has not been investigated. The objective of this study was to investigate the production of MIP-1alpha by basophils. Peripheral blood basophils were separated by Percoll gradient centrifugation, cultured overnight, and processed for double immunocytochemistry using Abs against MIP-1alpha and FcepsilonRIalpha (alpha subunit of IgE receptor type 1). We demonstrated that basophils expressed immunoreactive MIP-1alpha upon stimulation with anti-IgE. Less than 5% of the basophils stained for MIP-1alpha without stimulation. The secretion of MIP-1alpha by basophils was studied by ELISA. In these experiments, basophils were further enriched to 65 to 99% (median, 86%) by a negative selection method. Basophils released MIP-1alpha when stimulated by Abs against IgE and FCepsilonRIalpha as well as IL-3 and the calcium ionophore, A23187. In parallel experiments, PBMC, eosinophils, and neutrophils did not produce MIP-1alpha in response to anti-IgE, but they did so in response to A23187. No MIP-1alpha release was detected in platelet preparations. Preincubation with IL-3 (15 min or 18 h) augmented anti-IgE-included basophil MIP-1alpha production. The secretion of MIP-1alpha by basophils was detectable shortly after stimulation and gradually increased over 24 h. Since MIP-1alpha has potent inflammatory and histamine-releasing activities, its production by basophils may indicate a positive feedback mechanism for allergic inflammation.
Insights
This study reveals that basophils, a type of white blood cell, produce Macrophage inflammatory protein-1alpha (MIP-1alpha) when stimulated. This finding suggests a new role for basophils in allergic inflammation.
Area of Science:
- Immunology
- Cell Biology
- Allergy Research
Background:
- Macrophage inflammatory protein-1alpha (MIP-1alpha) is known to be produced by various immune cells.
- Production of MIP-1alpha by basophils, however, remained uninvestigated.
Purpose of the Study:
- To investigate the production of MIP-1alpha by human basophils.
- To explore the role of basophils in the context of allergic inflammation.
Main Methods:
- Isolation of peripheral blood basophils using Percoll gradient centrifugation and negative selection.
- Double immunocytochemistry to detect MIP-1alpha and FcεRIα expression.
- Enzyme-linked immunosorbent assay (ELISA) to quantify MIP-1alpha secretion.
- Stimulation of basophils with anti-IgE, anti-FcεRIα, IL-3, and calcium ionophore A23187.
Main Results:
- Basophils express and secrete MIP-1alpha upon stimulation with anti-IgE and anti-FcεRIα.
- IL-3 preincubation enhances anti-IgE-induced MIP-1alpha production by basophils.
- Unlike other myeloid cells, basophils produce MIP-1alpha in response to anti-IgE, but not A23187.
- MIP-1alpha secretion by basophils is a time-dependent process, increasing over 24 hours.
Conclusions:
- Basophils are a novel source of MIP-1alpha.
- Basophil-derived MIP-1alpha may contribute to allergic inflammation through a positive feedback mechanism.
- This discovery opens new avenues for understanding and potentially targeting allergic responses.
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