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• W1023733189 abstract "Abstract In a first set of experiments changes of intracellular free Ca2+ concentrations (Cai) within single cells were determined by flow cytometry in activated leukocytes loaded with the fluorescent Ca-indicator Indo-1. Stimulation of neutrophils or mononuclear cells with increasing amounts of FMLP led to a dose-dependent transient increase of Cai in neutrophils and monocytes, but not lymphocytes. Single neutrophils did not respond equally well to low or intermediate doses of FMLP (0.3nM or 1.0nM). Some were activated at earlier time points than others, whereas some were not activated at all. This differential response of single cells was seen with all the active ligands tested. Next ligands for Fc-receptors or complement receptor 1 (CR1) were tested in order to assess if crosslinking of these receptors results in changes of Cai. Incubation of neutrophils or mononuclear cells with crosslinking polyclonal rabbit anti-CR1 F(ab')2 did not result in an increase of Cai, whereas crosslinking of Fc-receptors with aggregated IgG or polyclonal rabbit anti-CR1 IgG did induce increases of Cai in monocytes and neutrophils, but not lymphocytes. Crosslinking of monoclonal anti-Fc-receptor F(ab')2 bound to neutrophils with F(ab')2 fragments of goat anti-mouse IgG led to an increase of Cai as well, proving that crosslinking of Fc-receptors is required for activation. Crosslinking with the second antibody of saturating amounts of monoclonal anti-CR1 F(ab')2 bound to neutrophils, instead of the monoclonal anti-Fc-receptor F(ab')2, did not result in any change of Cai. In a second set of experiments changes of CR3 expression on the surface of neutrophils and monocytes induced by the ligands for Fc-receptors and CR1 were measured. The ligands for Fc-receptors, aggregated IgG and polyclonal rabbit anti-CR1 IgG, induced dose-dependent upregulation of CR3 on neutrophils and monocytes. The same result was obtained upon crosslinking of monoclonal anti-Fc-receptor Fab' bound to neutrophils with F(ab')2 fragments of goat anti-mouse IgG. The expression of CR3 in neutrophils could not be induced by the CR1 crosslinking compounds, whereas polyclonal rabbit anti-CR1 F(ab')2 was able to induce slow CR3 upregulation in monocytes. But incubation of monocytes with monoclonal anti-CR1 Fab' followed by crosslinking of bound antibody with F(ab')2 goat anti-mouse IgG did not result in augmentation of CR3 expression. Finally, it was found that neutrophils depleted of intracellular Ca2+ still respond to activation by aggregated IgG with slow CR3 upregulation, showing that the increase of CR3 expression is not dependent on changes of Cai. It is concluded that crosslinking of Fc-receptors on neutrophils and monocytes, but not lymphocytes leads to changes of Cai and CR3 expression, which is not observed after CR1 crosslinking on neutrophils. Cai is not changed in monocytes as well upon CR1 crosslinking, whereas with one CR1 crosslinking ligand slow CR3 expression was induced showing that in these cells CR1 may be able to transmit a Cai independent signal into the interior leading to CR3 upregulation" @default.
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• W1023733189 date "1989-01-01" @default.
• W1023733189 modified "2023-10-18" @default.
• W1023733189 title "Measurement by Flow Cytometry of Intracellular Free Ca2+ Concentration and Complement Receptor Expression in Activated Neutrophils and Monocytes" @default.
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• W1023733189 doi "https://doi.org/10.1016/b978-0-08-037378-2.50041-3" @default.
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