Kinetics of GATA-1 expression in cord blood eosinophil progenitors: effects of Budesonide

Crawford, L., Baatjes, A.J., Cyr, M.M., Hayes, L.M. and Denburg, J.A. (2004) Kinetics of GATA-1 expression in cord blood eosinophil progenitors: effects of Budesonide. Journal of Allergy and Clinical Immunology, 113(2), S169. (doi: 10.1016/j.jaci.2004.01.042)

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<b>Rationale</b> The transcription factor GATA-1, has been shown to play a critical role in hemopoietic stem cell lineage commitment, especially to the eosinophilic pathway. However, little is known concerning the timing and responsiveness of GATA-1 expression in cytokine-induced eosinophil differentiation from cord blood (CB) progenitor cells. In this study we sought to determine the kinetics of GATA-1 expression after in-vitro stimulation of CB progenitors with interleukin-5 (IL-5), and the effects of a corticosteroid on this process.<p></p> <b>Methods</b> Non-adherent mononuclear cells (MNC) were isolated from CB, and incubated in the presence or absence of recombinant human IL-5 (1ng/ml) and/or Budesonide (10−8M). At various time-points post-stimulation, RNA was isolated and GATA-1 gene expression was determined after comparative Q-PCR. The fold increase between stimulated and un-stimulated cells was calculated using the 2−ΔΔCt method.<p></p> <b>Results</b> Stimulation with IL-5 resulted in marked up-regulation of GATA-1 expression between 24 and 72 hours, compared to baseline, with peak expression of approximately 18-fold relative to un-stimulated CBMNC at 72 hours. Preliminary data indicate that Budesonide treatment of IL-5 stimulated CBMNC resulted in an approximate 5-fold increase in GATA-1 expression at 4 hours, followed by suppression (10-fold decrease) at 24 hours, compared to the 4 hour time-point.<p></p> <b>Conclusions</b> In-vitro stimulation of CBMNC with IL-5 induces early and sustained GATA-1 up-regulation, which is later suppressed by a topically active corticosteroid. We are developing this method as a very sensitive, surrogate marker of eosinophil progenitor lineage commitment in ongoing studies of mechanisms of action of anti-allergic agents and in CB studies of the atopic diathesis.<p></p>

Item Type:Articles
Glasgow Author(s) Enlighten ID:Stewart, Ms Lynn
Authors: Crawford, L., Baatjes, A.J., Cyr, M.M., Hayes, L.M., and Denburg, J.A.
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Journal of Allergy and Clinical Immunology
Journal Abbr.:J. Allergy Clin. Immunol.
ISSN (Online):1097-6825

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