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Human cord blood derived immature basophils show dual characteristics, expressing both basophil and eosinophil associated proteins

Grundström, Jeanette and Reimer, Jenny M. and Magnusson, Sofia E. and Nilsson, Gunnar and Wernersson, Sara and Hellman, Lars (2012). Human cord blood derived immature basophils show dual characteristics, expressing both basophil and eosinophil associated proteins. PloS one. 7:10, 1-10
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Official URL: http://dx.doi.org/10.1371/journal.pone.0048308

Abstract

Basophils are blood cells of low abundance associated with allergy, inflammation and parasite infections. To study the
transcriptome of mature circulating basophils cells were purified from buffy coats by density gradient centrifugations and two-step magnetic cell sorting. However, after extensive analysis the cells were found to be transcriptionally inactive and almost completely lack functional mRNA. In order to obtain transcriptionally active immature basophils for analysis of their
transcriptome, umbilical cord blood cells were therefore cultured in the presence of interleukin (IL)-3 for 9 days and basophils were enriched by removing non-basophils using magnetic cell sorting. The majority of purified cells
demonstrated typical metachromatic staining with Alcian blue dye (95%) and expression of surface markers FceRI and
CD203c, indicating a pure population of cells with basophil-like phenotype. mRNA was extracted from these cells and used to construct a cDNA library with approximately 600 000 independent clones. This library served as tool to determine the mRNA frequencies for a number of hematopoietic marker proteins. It was shown that these cells express basophil/mast cellspecific transcripts, i.e. b-tryptase, serglycin and FceRI a-chain, to a relatively low degree. In contrast, the library contained a
high number of several eosinophil-associated transcripts such as: major basic protein (MBP), charcot leyden crystal (CLC), eosinophil cationic protein (ECP), eosinophil derived neurotoxin (EDN) and eosinophil peroxidase (EPO). Out of these transcripts, MBP and EPO were the most frequently observed, representing 8% and 3.2% of the total mRNA pool, respectively. Moreover, in a proteome analysis of cultured basophils we identified MBP and EPO as the two most prominent protein bands, suggesting a good correlation between protein and mRNA analyses of these cells. The mixed phenotype observed for these cells strengthens the conclusion that eosinophils and basophils are closely linked during human hematopoietic development. The dual phenotype also indicates that other cytokines than IL-3 or cell surface interactions are needed to obtain the full basophil specific phenotype in vivo.

Authors/Creators:Grundström, Jeanette and Reimer, Jenny M. and Magnusson, Sofia E. and Nilsson, Gunnar and Wernersson, Sara and Hellman, Lars
Title:Human cord blood derived immature basophils show dual characteristics, expressing both basophil and eosinophil associated proteins
Series/Journal:PloS one (1932-6203)
Year of publishing :October 2012
Volume:7
Number:10
Page range:1-10
Number of Pages:10
Publisher:Public Library of Science
ISSN:1932-6203
Language:English
Publication Type:Journal article
Refereed:Yes
Article category:Scientific peer reviewed
Version:Published version
Full Text Status:Public
Agris subject categories.:L Animal production > L50 Animal physiology and biochemistry
X Agricola extesions > X30 Life sciences
Subjects:(A) Swedish standard research categories 2011 > 3 Medical and Health Sciences > 301 Basic Medicine > Cell and Molecular Biology
Keywords:eosinophils , basophils, blood cells, mRNA, cDNA library, proteins, hematopoietic development, dual phenotype
URN:NBN:urn:nbn:se:slu:epsilon-e-1651
Permanent URL:
http://urn.kb.se/resolve?urn=urn:nbn:se:slu:epsilon-e-1651
Additional ID:
Type of IDID
Web of Science (WoS)000310600500098
DOI10.1371/journal.pone.0048308
ID Code:10838
Department:(VH) > Dept. of Anatomy, Physiology and Biochemistry
Deposited By: SLUpub Connector
Deposited On:04 Oct 2013 11:26
Metadata Last Modified:01 Apr 2015 10:38

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