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Mechanisms in Neutrophil Priming: Characterization of the Oxidative Response Induced by Formylmethionyl‐Leucyl‐Phenylalanine in Human Exudated Cells
Author(s) -
FOLLIN P.,
BRIHEIM G.,
DAHLGREN C.
Publication year - 1991
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1111/j.1365-3083.1991.tb01552.x
Subject(s) - ionomycin , exudate , cytochalasin b , diacylglycerol kinase , priming (agriculture) , ionophore , chemistry , respiratory burst , cytochalasin , concanavalin a , incubation , in vitro , biochemistry , stimulation , biology , endocrinology , enzyme , cell , protein kinase c , membrane , cytoskeleton , botany , germination
Exudate human polymorphonuclear neutrophils were isolated and investigated regarding oxidative responsiveness and priming ability. The exudate neutrophils were found to produce an increased amount of O 2 ‐ and H 2 O 2 when stimulated with formylmethionyl‐leucyl‐phenylalanine (fMLP). i.e. these cells were metabolically primed. Cytochalasin B (cyl B) pretreatment affected the production of O 2 ‐ by exudate cells, although to a lesser extent than the production by peripheral blood cells, in which a substantial increase was induced. Addition of N‐ethylmaleimide (NEM) to activated exudate and peripheral blood cells revealed no difference in oxidase in activation rate. To induce further priming, the cells were incubated in vitro with a synthetic diacylglycerol (sn‐1,2‐didecanoylglycerol: diC 10 ), or the Ca 2+ ionophore ionomycin. Results of this procedure showed significant differences between exudate and peripheral blood neutrophils: the peripheral cells expressed a primed response, which was measured as increased fMLP‐induced O 2 ‐ production following incubation with both these substance: where a the metabolic activity of exudated cells was not affected by diC 10 but was significantly primed by ionomycin (P<0.01). The exact route for diacylglycerol priming is unknown. However, our results with human neutrophils primed during exudation indicate an exhausted diC 10 ‐priming pathway, with a retained sensitivity for priming [Ca 2+ ] i rises.