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Cord blood molecular biomarkers of eosinophilopoiesis: Kinetic analysis of GATA‐1, MBP1 and IL‐5Rα mRNA expression
Author(s) -
Ellis Anne K.,
Ackerman Steven J.,
Crawford Lynn,
Du Jian,
Bedi Richa,
Denburg Judah A.
Publication year - 2010
Publication title -
pediatric allergy and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.269
H-Index - 89
eISSN - 1399-3038
pISSN - 0905-6157
DOI - 10.1111/j.1399-3038.2010.01003.x
Subject(s) - medicine , cord blood , messenger rna , expression (computer science) , immunology , biochemistry , gene , biology , computer science , programming language
Ellis AK, Ackerman SJ, Crawford L, Du J, Bedi R, Denburg JA. Cord blood molecular biomarkers of eosinophilopoiesis: Kinetic analysis of GATA‐1, MBP1 and IL‐5Rα mRNA expression.
Pediatr Allergy Immunol 2010: 21: 640–648.
© 2010 John Wiley & Sons A/S Eosinophil/basophil (Eo/B) progenitor phenotype and function in cord blood (CB) are associated with atopic risk at birth and infant clinical outcomes. Molecular analyses of eosinophil–basophil differentiation events could identify clinically predictive biomarkers. To determine CB kinetic patterns of Eo/B lineage‐associated gene expression (GATA‐1, MBP1 and IL‐5Rα) after IL‐5 stimulation, CB non‐adherent mononuclear cells were isolated from random fresh and frozen samples and incubated in the presence of recombinant human interleukin‐5. Some underwent CD34+ positive selection using magnetic cell separation. At various time‐points, mRNA expression of GATA‐1, MBP1 and IL‐5Rα (total transcripts) was determined utilizing multiplex quantitative polymerase chain reaction (Q‐PCR). Relative expression levels of the IL‐5Rα soluble vs. transmembrane isoforms were also analyzed. Stimulation of the non‐adherent mononuclear cells with IL‐5 resulted in early up‐regulation of GATA‐1, peaking at 48 h, followed by decreasing expression and down‐regulation by 96 h. The CD34+ enriched population demonstrated an equivalent expression pattern ( r = 0.963, p = 0.0349). MBP1 mRNA expression [non‐adherent mononuclear cells (NAMNCs) and CD34+ alike; r = 0.988, p = 0.012] was slowly up‐regulated in response to IL‐5, maximal at 96 h. Total IL‐5Rα expression appeared stable over the time‐course, mediated by differential expression of the soluble and transmembrane isoforms (i.e., initial increase in the transmembrane contribution followed by a predominance of the soluble isoform by 48–72 h). Multiplex Q‐PCR analysis of mRNA from CB demonstrates expression of critical eosinophil‐basophil lineage‐specific events that are consistent with current understanding of eosinophil differentiation and maturation. The non‐adherent mononuclear cell population provides a surrogate signal for the CD34+ progenitor population.