PMID- 3817866
OWN - NLM
STAT- MEDLINE
DCOM- 19870406
LR  - 20181113
IS  - 0019-2805 (Print)
IS  - 1365-2567 (Electronic)
IS  - 0019-2805 (Linking)
VI  - 60
IP  - 1
DP  - 1987 Jan
TI  - Purification of eosinophils from normal human blood, preparation of eosinoplasts 
      and characterization of their functional response to various stimuli.
PG  - 123-9
AB  - Eosinophils from the blood of normal individuals were purified by centrifugation 
      over discontinuous Percoll gradients. Eosinophil suspensions were obtained with a 
      mean purity of 96% and a mean recovery of 64% (n = 19). When incubated with 
      phorbol-myristate acetate, eosinophils consumed twice as much oxygen as did 
      neutrophils from the same donors. With serum-treated zymosan, 70% and 100% of the 
      maximal oxidative response (i.e. the response to phorbol-myristate acetate) was 
      obtained with eosinophils and neutrophils, respectively. The calcium ionophore 
      A23187 is a weak stimulus that triggered only 2.5% of the eosinophil and 10% of 
      the neutrophil oxidative capacity. The response of both cell types to 
      formyl-methionyl-leucyl-phenylalanine (fMLP) was rapid, with a maximum after 3 
      min. The magnitude of this eosinophil reaction was half that of neutrophils. 
      Although the activities of the granule enzymes beta-glucuronidase and 
      arylsulphatase were 2.5 and 6 times higher in eosinophils than in neutrophils, 
      respectively, the exocytosis of these enzymes in response to various stimuli was 
      lower in eosinophils. The high yield of eosinophils from our separation method 
      enabled us to prepare eosinoplasts by centrifugation of eosinophils over 
      discontinuous Ficoll gradients that contained cytochalasin B. Eosinoplasts are 
      plasma membrane vesicles derived from eosinophils, filled with cytoplasm but 
      devoid of granules and nucleus. The eosinoplasts contained 30% of the cytoplasm 
      and plasma membrane present in intact eosinophils. Eosinoplasts still possessed a 
      functionally intact oxidase enzyme that could be stimulated with various stimuli. 
      Therefore, eosinoplasts may provide a valuable tool to study separately the role 
      of the oxidase products and that of the granule contents in eosinophil functions.
FAU - Yazdanbakhsh, M
AU  - Yazdanbakhsh M
FAU - Eckmann, C M
AU  - Eckmann CM
FAU - De Boer, M
AU  - De Boer M
FAU - Roos, D
AU  - Roos D
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Immunology
JT  - Immunology
JID - 0374672
RN  - 0 (Cytochrome b Group)
RN  - 9064-78-2 (cytochrome b558)
RN  - EC 1.6.3.- (NADPH Oxidases)
RN  - EC 3.1.6.1 (Arylsulfatases)
RN  - EC 3.2.1.31 (Glucuronidase)
RN  - S88TT14065 (Oxygen)
SB  - IM
MH  - Arylsulfatases/metabolism
MH  - Cell Separation/*methods
MH  - Cell Survival
MH  - Centrifugation, Density Gradient
MH  - Cytochrome b Group/analysis
MH  - Cytoplasm/metabolism
MH  - Eosinophils/cytology/enzymology/*metabolism
MH  - Glucuronidase/metabolism
MH  - Humans
MH  - Luminescent Measurements
MH  - *NADPH Oxidases
MH  - Oxygen/metabolism
PMC - PMC1453348
EDAT- 1987/01/01 00:00
MHDA- 1987/01/01 00:01
PMCR- 1988/01/01
CRDT- 1987/01/01 00:00
PHST- 1987/01/01 00:00 [pubmed]
PHST- 1987/01/01 00:01 [medline]
PHST- 1987/01/01 00:00 [entrez]
PHST- 1988/01/01 00:00 [pmc-release]
PST - ppublish
SO  - Immunology. 1987 Jan;60(1):123-9.