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The Capital Region of Denmark - a part of Copenhagen University Hospital
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Expression of the Transcription Factor E4BP4 in Human Basophils

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Rationale
The cytokine IL-3 plays an important role for human basophil development, function and survival.
IL-3 is also reported to induce the expression of the transcription factor E4BP4, but it is not known whether E4BP4 is expressed in basophils and influences basophil responsiveness. The aim of this study was to determine whether human basophils express E4BP4 and if so, to compare the expression of E4BP4 with basophil histamine release.

Methods
Buffy coat blood (<6h) was analyzed for anti-IgE mediated histamine release. Basophils were purified by negative selection and purity was determined by Alcian blue. RNA was extracted (0.005-0.02 µg RNA from 0.5 - 1 x 106 cells), and the corresponding cDNA analyzed by real-time PCR where E4BP4 expression was calculated as 2-(CT(E4BP4) - CT(β-actin)). E4BP4 protein expression was visualized in basophil lysates (107 cells/ml) by Western blot followed by ECL stain and X-ray film exposure. Protein band intensities were correlated to -actin expression.

Results
We analyzed basophils from 14 donors and found E4BP4 mRNA expression in all donors (2.33 ± 2.42) despite a low basophil RNA level. Seven donors were also tested for E4BP4 protein expression and again all basophil preparations were found positive.
Comparing the expression of E4BP4 with anti-IgE mediated histamine release revealed a trend towards a positive correlation between E4BP4 protein expression and basophil releasability (r = 0.37, p = 0.42).

Conclusion
Human basophils express the transcription factor E4BP4 which might have an impact on basophil histamine release.
Original languageEnglish
Publication date2014
Publication statusPublished - 2014

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