A new and simple method for studying the binding and ingestion steps in the phagocytosis of yeasts
Identifieur interne : 002E43 ( Main/Exploration ); précédent : 002E42; suivant : 002E44A new and simple method for studying the binding and ingestion steps in the phagocytosis of yeasts
Auteurs : J. Giaimis [France] ; Y. Lombard [France] ; M. Makaya-Kumba [France] ; P. Fonteneau [France] ; P. Poindron [France]Source :
- Journal of Immunological Methods [ 0022-1759 ] ; 1992.
English descriptors
- Teeft :
- Academic press, Acta pathol, Alternative pathway, Autoclaved, Autoclaved yeasts, Biol, Blood clearance, Cell biol, Chloroquine, Culture medium, Cytochalasin, Deep violet color, Escherichia coil, Escherichia coli, Extracellular yeasts, Fetal calf serum, Flow cytometry, Identical experiments, Ingestion, Internalization steps, Intracellular, Intracellular yeasts, Leukocyte biol, Light microscopy, Macrophage, Macrophage membrane, Malu, Malu cells, Malu cells phagocytosing autoclaved yeasts, Mannose, Mannose receptors, Microbial surface, Microorganism, Mononuclear phagocytes, Ofek, Optimal ratio, Phagocytosing cells, Phagocytosis, Phagocytosis test, Pink yeast, Pink yeasts, Receptor, Receptor recycling, Simple method, Staining properties, Surface lectins, Tannic, Tannic acid, Tannic acid solution, Tannic acid treatment, Total number, Total yeasts, Violet yeasts, Yeast.
Abstract
Abstract: Autoclaved yeasts are stained light pink by May-Grünwald Giemsa (MGG). If treated with tannic acid solution just before MGG staining, they display a deep violet color. It seemed possible that these properties could be used to discriminate between extra- and intracellular yeasts in a phagocytosis test, extracellular yeasts being violet and intracellular yeasts being pink. To validate this protocol, quantitative studies of phagocytosis by MALU cells (a murine macrophage cell line) were performed in the presence or absence of drugs known to interfere with phagocytosis.After treatment of cells with cytochalasin B, the mean number of pink yeasts per cell decreased in a dose-dependent manner, the mean number of violet yeasts increased in a dose-dependent manner, whereas the total number of cell-associated yeasts remained almost unchanged whatever the dose used. After treatment with α-mannans or chloroquine, the mean numbers of both violet and pink yeasts decreased in a dose-dependent manner.These results confirmed that (i) violet yeasts are extracellular, (ii) autoclaved yeasts recognize lectin receptors, and (iii) unstained (pink) yeasts are intracellular. We show that this simple method can be used for quantitative light microscopic analysis of both the attachment and internalization steps in the phagocytosis of yeasts.
Url:
DOI: 10.1016/0022-1759(92)90191-U
Affiliations:
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<term>Autoclaved yeasts</term>
<term>Biol</term>
<term>Blood clearance</term>
<term>Cell biol</term>
<term>Chloroquine</term>
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<term>Cytochalasin</term>
<term>Deep violet color</term>
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<term>Escherichia coli</term>
<term>Extracellular yeasts</term>
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<term>Flow cytometry</term>
<term>Identical experiments</term>
<term>Ingestion</term>
<term>Internalization steps</term>
<term>Intracellular</term>
<term>Intracellular yeasts</term>
<term>Leukocyte biol</term>
<term>Light microscopy</term>
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<term>Macrophage membrane</term>
<term>Malu</term>
<term>Malu cells</term>
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<term>Mannose receptors</term>
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<term>Mononuclear phagocytes</term>
<term>Ofek</term>
<term>Optimal ratio</term>
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<term>Phagocytosis</term>
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<term>Receptor recycling</term>
<term>Simple method</term>
<term>Staining properties</term>
<term>Surface lectins</term>
<term>Tannic</term>
<term>Tannic acid</term>
<term>Tannic acid solution</term>
<term>Tannic acid treatment</term>
<term>Total number</term>
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<front><div type="abstract" xml:lang="en">Abstract: Autoclaved yeasts are stained light pink by May-Grünwald Giemsa (MGG). If treated with tannic acid solution just before MGG staining, they display a deep violet color. It seemed possible that these properties could be used to discriminate between extra- and intracellular yeasts in a phagocytosis test, extracellular yeasts being violet and intracellular yeasts being pink. To validate this protocol, quantitative studies of phagocytosis by MALU cells (a murine macrophage cell line) were performed in the presence or absence of drugs known to interfere with phagocytosis.After treatment of cells with cytochalasin B, the mean number of pink yeasts per cell decreased in a dose-dependent manner, the mean number of violet yeasts increased in a dose-dependent manner, whereas the total number of cell-associated yeasts remained almost unchanged whatever the dose used. After treatment with α-mannans or chloroquine, the mean numbers of both violet and pink yeasts decreased in a dose-dependent manner.These results confirmed that (i) violet yeasts are extracellular, (ii) autoclaved yeasts recognize lectin receptors, and (iii) unstained (pink) yeasts are intracellular. We show that this simple method can be used for quantitative light microscopic analysis of both the attachment and internalization steps in the phagocytosis of yeasts.</div>
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