Hypoxia modulates human eosinophil function.

Clinical and Molecular Allergy : CMA
Alon H Nissim Ben EfraimF Levi-Schaffer

Abstract

Eosinophils are involved in various inflammatory processes including allergic inflammation during which angiogenesis has been documented. Angiogenesis is most likely connected to the hypoxia which characterizes inflamed tissues. Eosinophils produce VEGF and are pro-angiogenic. However, to the best of our knowledge no study has been performed to verify the existence of a direct link between eosinophils, hypoxia and angiogenesis in allergic inflammation. To characterize eosinophil function and angiogenic potential under hypoxic conditions. Human peripheral blood eosinophils were cultured in normoxic or hypoxic conditions with or without cytokines. Viability and apoptosis were assessed by Annexin V/PI staining. Anti- or pro-apoptotic protein levels, HIF-1alpha levels and MAPK phosphorylation were analyzed by immunoblot analysis. Angiogenic mediator release was evaluated by ELISA. Hypoxic eosinophils were more viable than normoxic ones after up to three days. In addition in hypoxia, anti-apoptotic Bcl-XL protein levels increased more than pro-apoptotic Bax levels. Hypoxia increased VEGF and IL-8 release. In hypoxic eosinophils high levels of HIF-1alpha were observed, particularly in the presence of GM-CSF. MAPK, particularly ERK1/2...Continue Reading

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Methods Mentioned

BETA
flow cytometry
ELISA
ELISAs

Software Mentioned

CellQuest
[UNK]

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