A new and simple method for studying the binding and ingestion steps in the phagocytosis of yeasts

Th1 and Th2 cytokines cooperate to stimulate mannose-receptor-mediated phagocytosis

“…Cells were incubated with cytokines in tissue culture medium for varying lengths of time before the addition of yeast. Yeast were added at a ratio of 200:1 macrophage in ␣-MEM without serum at 37°C for 20 min before fixation with 1% tannic acid [17]. Internalized yeast were assayed by the differential staining method of Giaimis et al [17].…”

“…Yeast were added at a ratio of 200:1 macrophage in ␣-MEM without serum at 37°C for 20 min before fixation with 1% tannic acid [17]. Internalized yeast were assayed by the differential staining method of Giaimis et al [17]. This protocol stains extracellular yeast dark blue and internalized yeast light pink.…”

“…The cells were then incubated for 20 min at 37°C with heat-killed Saccharomyces cerevesiae. Nonadherent particles were removed and the cells were differentially stained in order to assess ingestion of yeast [17].…”

Noninfectious entry of HIV-1 into peripheral and brain macrophages mediated by the mannose receptor

“…Binding of HIV-1 gp120 to the Macrophage MR. To determine whether HIV-1 gp120 binds to the MR of macrophages, an MR-mediated phagocytosis assay of yeast particles using murine alveolar macro-phages was performed, as described (32). The extent to which recombinant HIV-1 gp120 (rgp120) blocked this pathway was observed.…”

“…Human and mouse alveolar macrophages and human microglia were used to ingest yeast particles and fluoresceinated PC, as described (see SI Materials and Methods) (27,45). The phagocytosis assay was performed by using the technique of Giaimis et al (32).…”