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The present report describes the increase in cytosolic‐free calcium levels (Ca2+i) induced by chemoattractants in polymorphonuclear leukocytes (PMN) using the new fluorescent Ca2+ chelator Indo‐1 AM. Increases in cytosolic Ca2+ were measured by flow cytometry. With this approach, 98% of PMN were found to respond to F‐Met‐Leu‐Phe (FMLP) and leukotriene B4 (LTB4) stimulation. Although both substances induced a rapid and relatively homogeneous rise in Ca2+i, only FMLP gave a sustained Ca2+i rise, whereas that induced by LTB4 appeared transient. In addition, the combined use of wide‐angle light scatter and electronic cell volume measurement allowed analysis of Ca2+i variations in the total peripheral blood cell population. The supernatant of a human fibrous histiocytoma cell line (GCT) was able to increase Ca2+ release in PMN. This activity may be ascribed to a new granulocytic activation factor, as neither human recombinant interleukin‐1α (IL‐1α), granulocyte‐colony stimulating factor (G‐CSF), nor granulocyte‐macrophage‐colony stimulating factor (GM‐CSF), which were present in this supernatant, were able to induce a Ca2+i rise in PMN.
Cytometry Part A – Wiley
Published: Mar 1, 1989
Keywords: ; ; ; ; ;
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