Kinetics of cord blood eosinophil lineage biomarker mRNA: co-expression of CysLTR1 with GATA-1

Crawford, L., Ellis, A.K. and Denburg, J.A. (2007) Kinetics of cord blood eosinophil lineage biomarker mRNA: co-expression of CysLTR1 with GATA-1. Journal of Allergy and Clinical Immunology, 119(1), S215. (doi: 10.1016/j.jaci.2006.12.213)

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<b>RATIONALE:</b> We and others have shown that hemopoietic progenitors express CysLT receptors. Our laboratory has also shown that, upon stimulation with IL-5, cord blood mononuclear cells (CB MNC's) demonstrate kinetic mRNA expression patterns of key eosinophil-lineage specific genes, namely GATA-1 and MBP, which are concordant with current understanding of eosinophil developmental biology. Our objective was to evaluate CysLTR1 mRNA expression by Q-PCR analysis, and assess changes and determine the kinetic relationship of CysLTR1 to GATA-1 mRNA expression in CB MNC's after in-vitrostimulation with IL-5. <p></p> <b>METHODS:</b> Non-adherent CB MNCs were isolated from random fresh and frozen samples, and incubated in the presence of IL-5 (1 ng/mL). At 24, 48 and 72h post-stimulation, RNA was isolated, reverse transcribed, and expression of GATA-1 and CysLTR1 was determined utilizing multiplex Q-PCR. Relative expression was calculated using the delta-delta Ct method. GATA-1 and CysLTR1 expression were compared using Pearson's correlation.<p></p> <b>RESULTS:</b> Stimulation with IL-5 resulted in: 1) up-regulation of CysLTR1 mRNA expression, peaking between 24-48hrs, with a fall by 72hrs; 2) similar, but more robust, up-regulation of GATA-1 mRNA expression, also peaking between 24-48hrs, with slight down-regulation by 72hr. The expression patterns for CysLTR1 and GATA-1 were highly correlated with each other (overall R2 = 0.993).<p></p> <b>CONCLUSIONS:</b> Multiplex Q-PCR analysis of mRNA from CB MNC's stimulated with IL-5 demonstrates changes in the expression of CysLTR1 in a pattern which mirrors that of GATA-1, a critical eosinophil-lineage transcription factor. The close co-expression of these markers suggests that early eosinophil-lineage commitment may be functionally influenced by CysLTs.<p></p>

Item Type:Articles
Glasgow Author(s) Enlighten ID:Stewart, Ms Lynn
Authors: Crawford, L., Ellis, A.K., 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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