Updates on the surface antigens of basophils: CD16 on basophils of patients with respiratory or insect venom allergy and the rejection of CD203c and CD63 externalization decoupling by bisindolylmaleimides
Language English Country England, Great Britain Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
30288810
DOI
10.1111/cea.13288
Knihovny.cz E-resources
- Keywords
- FcγRIIIA, IgG-mediated anaphylaxis, basophil activation test, bisindolylmaleimide, flow cytometry artifact,
- MeSH
- Hypersensitivity immunology pathology MeSH
- Tetraspanin 30 immunology MeSH
- Basophils immunology pathology MeSH
- Adult MeSH
- Phosphoric Diester Hydrolases immunology MeSH
- GPI-Linked Proteins immunology MeSH
- Indoles chemistry MeSH
- Arthropod Venoms toxicity MeSH
- Insect Bites and Stings immunology pathology MeSH
- Middle Aged MeSH
- Humans MeSH
- Maleimides chemistry MeSH
- Pyrophosphatases immunology MeSH
- Receptors, IgG immunology MeSH
- Aged MeSH
- Check Tag
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Aged MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Tetraspanin 30 MeSH
- bisindolylmaleimide MeSH Browser
- CD63 protein, human MeSH Browser
- ENPP3 protein, human MeSH Browser
- FCGR3B protein, human MeSH Browser
- Phosphoric Diester Hydrolases MeSH
- GPI-Linked Proteins MeSH
- Indoles MeSH
- Arthropod Venoms MeSH
- Maleimides MeSH
- Pyrophosphatases MeSH
- Receptors, IgG MeSH
BACKGROUND: CD16 was previously suggested to be a new marker of basophils that is subject to downregulation by FcεRI crosslinking. Certain compounds, including supraoptimal concentrations of the PKC inhibitors, bisindolylmaleimides, decouple the release of granules containing CD203c, CD63 and histamine, and may thus help to identify the mechanisms related to the CD16 externalization. OBJECTIVE: We hypothesized that CD16 is differentially expressed on the surface of basophils in patients with birch pollen or insect venom allergy and is subject to a regulation in response to allergens. We also employed CD203c and CD63 externalization decoupling by bisindolylmaleimides. METHODS: We performed a basophil activation test coupled with CD16 and histamine detection using cells isolated from patients with allergy to birch pollen or insect venom and negative controls. We employed two PKC inhibitors, bisindolylmaleimide II and Ro 31-8220 at their supraoptimal concentrations and, after difficulties reproducing previously published data, we analyzed the fluorescence of these inhibitors alone. We identified the CD16 isoforms by sequencing nested RT-PCR amplicons from flow cytometry sorted basophils and by cleaving the CD16b GPI anchor using a phospholipase C. RESULTS: We provide the first evidence that CD16a is expressed as a surface antigen on a small subpopulation of human basophils in patients with respiratory and insect venom allergy, and this antigen shows increased surface expression following allergen challenge or FcεRI crosslinking. We rejected the apparent decoupling of the surface expression of basophil activation markers following the administration of bisindolylmaleimides. CONCLUSIONS & CLINICAL RELEVANCE: The inclusion of αCD16 in negative selection cocktails selects against a subset of basophils that are CD16+ or CD16dim . Using CD16dim basophils and unstained leucocytes, we show that previous studies with supraoptimal concentrations of bisindolylmaleimides are likely flawed and are not associated with the differential expression of CD203c and CD63.
2nd Department of Internal Medicine 3rd Faculty of Medicine Charles University Prague Czech Republic
Department of Immunology 3rd Faculty of Medicine Charles University Prague Czech Republic
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