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      Role of calcium during lipopolysaccharide stimulation of neutrophils.

      Infection and Immunity
      Antigens, CD18, metabolism, Calcium, Chelating Agents, pharmacology, Egtazic Acid, Fluorescent Dyes, Fura-2, Humans, In Vitro Techniques, Lipopolysaccharides, Macrophage-1 Antigen, Models, Biological, N-Formylmethionine Leucyl-Phenylalanine, Neutrophils, drug effects, immunology, Second Messenger Systems, Spectrometry, Fluorescence, Terpenes, Thapsigargin

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          Abstract

          This study investigated the role of intracellular calcium concentration ([Ca]i) as a possible intermediate in the lipopolysaccharide (LPS) second messenger pathway for the activation of neutrophils (polymorphonuclear leukocytes [PMNs]). Isolated PMNs were loaded with the calcium-sensitive fluorescent dye fura-2. The PMNs were stimulated with either LPS or the positive control formyl-Met-Leu-Phe (fMLP). As expected, PMN exposure to fMLP increased [Ca]i. However, LPS stimulation did not induce any detectable changes. Depletion of intracellular Ca stores with thapsigargin, or extracellular Ca with EGTA, significantly inhibited the upregulation of the CD11b/CD18 integrin in response to fMLP but not LPS. We conclude that [Ca]i is not an early intermediate in the second-messenger pathway for the activation of PMNs by LPS.

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