Analysis of cytosolic ionized calcium variation in polymorphonuclear leukocytes using flow cytometry and Indo‐1 AM

Lopez, Marc; Olive, Daniel; Mannoni, P

doi:10.1002/cyto.990100207

Cited by 20 publications

(5 citation statements)

References 32 publications

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“…Intracellular concentrations of Ca 2ϩ were measured using indo-1/AM according to a previously described method (58). Purified monocytes were resuspended in HBSS containing 1 mM Ca 2ϩ , 1 mM Mg 2ϩ , and 5% autologous serum and incubated with 10 mM indo-1/AM for 45 min at 37°C.…”

Section: Measurement Of Ca 2ϩ Influx In Monocytesmentioning

confidence: 99%

Human Galectin-3 Is a Novel Chemoattractant for Monocytes and Macrophages

Sano

Hsu

et al. 2000

The Journal of Immunology

468

344

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Galectin-3 is a β-galactoside-binding protein implicated in diverse biological processes. We found that galectin-3 induced human monocyte migration in vitro in a dose-dependent manner, and it was chemotactic at high concentrations (1.0 μM) but chemokinetic at low concentrations (10–100 nM). Galectin-3-induced monocyte migration was inhibited by its specific mAb and was blocked by lactose and a C-terminal domain fragment of the protein, indicating that both the N-terminal and C-terminal domains of galectin-3 are involved in this activity. Pertussis toxin (PTX) almost completely blocked monocyte migration induced by high concentrations of galectin-3. Galectin-3 caused a Ca2+ influx in monocytes at high, but not low, concentrations, and both lactose and PTX inhibited this response. There was no cross-desensitization between galectin-3 and any of the monocyte-reactive chemokines examined, including monocyte chemotactic protein-1, macrophage inflammatory protein-1α, and stromal cell-derived factor-1α. Cultured human macrophages and alveolar macrophages also migrated toward galectin-3, but not monocyte chemotactic protein-1. Finally, galectin-3 was found to cause monocyte accumulation in vivo in mouse air pouches. These results indicate that galectin-3 is a novel chemoattractant for monocytes and macrophages and suggest that the effect is mediated at least in part through a PTX-sensitive (G protein-coupled) pathway.

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Section: Measurement Of Ca 2ϩ Influx In Monocytesmentioning

confidence: 99%

Human Galectin-3 Is a Novel Chemoattractant for Monocytes and Macrophages

Sano

Hsu

et al. 2000

The Journal of Immunology

468

344

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“…Cells in suspension ( 5 , 16,35) or attached to a supporting matrix (31,32) can be analyzed as a population by spectrofluorometry. For single cell analysis, cells in suspension can be studied by flow cytometry (10,29), while attached cells are more amenable for study by image cytometry (24,32).…”

Section: Introductionmentioning

confidence: 99%

Attachment of A172 human glioblastoma cells affects calcium signalling: A comparison of image cytometry, flow cytometry, and spectrofluorometry

Szöllősi¹,

Feuerstein²,

Hyun³

et al. 1991

Cytometry

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The intracellular free calcium concentration ([Ca2'li) of indo-1 loaded A172 human glioblastoma cells stimulated by platelet-derived growth factor (PDGF) was studied in cell suspensions by flow cytometry and spectrofluorometry and in confluent monolayers by laser image cytometry and spectrofluorometry. With all three techniques, the percentage of responsive cells, peak [Ca2+li, and the duration of response were directly related, and the delay time was inversely related to PDGF dose. The maximum response occurred at a PDGF concentration of about 20 nglml. Basal and peak [Ca2+li did not differ significantly from method to method even though different calibration procedures were used. Cells in suspension monitored by both spectrofluorometry and flow cytometry displayed significantly shorter calcium responses than attached cells. This did not appear to be a direct effect of trypsinization. Spectral analysis of indo-1 in cytoplasm, 40% glycerol, and aqueous solutions showed significant differences in the isosbestic point and quantum efficiency. Calibration of [Ca2+li with spectrofluorometry is more accurate using the ratio of fluorescence intensities than the fluorescence intensities measured at either 405 or 485 nm.

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“…ing changes in cytosolic calcium levels and heterogeneity in cell responses to stimulators (4,8,9,15,19,27). air-cooled lasers, which emit only a t 488 nm have become popular because of their ease of maintenance and lower cost.…”

Section: Flow Cytometric Methods Have Been Useful In Study-mentioning

confidence: 99%