PMID- 16918508
OWN - NLM
STAT- MEDLINE
DCOM- 20070110
LR  - 20081121
IS  - 0105-4538 (Print)
IS  - 0105-4538 (Linking)
VI  - 61
IP  - 9
DP  - 2006 Sep
TI  - Isolation of transcriptionally active umbilical cord blood-derived basophils 
      expressing Fc epsilon RI, HLA-DR and CD203c.
PG  - 1063-70
AB  - BACKGROUND: Basophils are inflammatory cells associated with allergy and parasite 
      infections. Investigation of their true biological function has long been 
      hampered by the difficulty in obtaining sufficient amounts of pure basophils and 
      by the lack of phenotypic markers. Moreover, it has been very difficult to clone 
      and identify basophil-specific granule proteins, partially because of an almost 
      complete lack of mRNA in mature circulating basophils. METHODS: To obtain 
      transcriptionally active immature basophils, umbilical cord blood cells were 
      cultured in the presence of interleukin (IL)-3. The cells were analysed by flow 
      cytometry and by histological staining. RESULTS: The continuous presence of IL-3 
      in cord blood cultures resulted in the expansion of basophil precursors 
      co-expressing FcepsilonRI and the recently described mast cell/basophil marker, 
      97A6 (CD203c). Several nonbasophil markers (i.e. CD3, CD14, CD15, CD16, CD19 and 
      CD21) were absent on the cultured basophils. However, we show that in early 
      cultures, almost 60% of the CD203c+ cells co-express human leukocyte antigen 
      (HLA)-DR, a marker that is absent on mature circulating basophils. The presence 
      of HLA-DR on basophil precursors may explain the low recovery (24+/-5.2%) 
      obtained after isolation of cultured basophils, when using a conventional 
      basophil isolation kit that removes HLA-DR+ cells. A novel purification method 
      was developed, including a two-step cocktail of antibodies against selected 
      markers, which resulted in both high purity (95+/-0.5%) and recovery (59+/-1.5%) 
      of cultured basophils. CONCLUSIONS: We here establish cord blood cultures as a 
      source from which transcriptionally active basophil precursors can be isolated in 
      reasonable quantities for thorough biochemical characterization.
FAU - Reimer, J M
AU  - Reimer JM
AD  - Department of Cell and Molecular Biology, Uppsala University, Uppsala, Sweden.
FAU - Magnusson, S
AU  - Magnusson S
FAU - Juremalm, M
AU  - Juremalm M
FAU - Nilsson, G
AU  - Nilsson G
FAU - Hellman, L
AU  - Hellman L
FAU - Wernersson, S
AU  - Wernersson S
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Denmark
TA  - Allergy
JT  - Allergy
JID - 7804028
RN  - 0 (ENPP3 protein, human)
RN  - 0 (HLA-DR Antigens)
RN  - 0 (Receptors, IgE)
RN  - EC 3.1.4.- (Phosphoric Diester Hydrolases)
RN  - EC 3.6.1.- (Pyrophosphatases)
SB  - IM
CIN - Allergy. 2006 Sep;61(9):1025-7. PMID: 16918503
MH  - Basophils/*immunology/*metabolism
MH  - Cell Separation
MH  - Cells, Cultured
MH  - Fetal Blood/*cytology/*immunology/metabolism
MH  - HLA-DR Antigens/biosynthesis/*genetics
MH  - Humans
MH  - Phosphoric Diester Hydrolases/biosynthesis/*genetics
MH  - Pyrophosphatases/biosynthesis/*genetics
MH  - Receptors, IgE/*biosynthesis/*genetics
MH  - Stem Cells/immunology/metabolism
MH  - Transcriptional Activation/immunology
MH  - Umbilical Veins
EDAT- 2006/08/22 09:00
MHDA- 2007/01/11 09:00
CRDT- 2006/08/22 09:00
PHST- 2006/08/22 09:00 [pubmed]
PHST- 2007/01/11 09:00 [medline]
PHST- 2006/08/22 09:00 [entrez]
AID - ALL1149 [pii]
AID - 10.1111/j.1398-9995.2006.01149.x [doi]
PST - ppublish
SO  - Allergy. 2006 Sep;61(9):1063-70. doi: 10.1111/j.1398-9995.2006.01149.x.