Flow cytometric quantitation of attachment and phagocytosis in phenotypically-defined subpopulations of cells using PKH26-labeled Fc γR-specific probes

Human receptors for IgG (Fc γR) are characterized by diverse structure and function. We describe a flow cytometric technique to quantitate receptor-specific Fc γR-mediated attachment and phagocytosis in phenotypically-defined subsets of cells using erythrocytes (E) labeled with the lipophilic dye PK...

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Bibliographic Details
Published in:Journal of immunological methods 1997-06, Vol.205 (1), p.55-65
Main Authors: Pricop, Luminita, Salmon, Jane E, Edberg, Jeffrey C, Beavis, Andrew J
Format: Article
Language:English
Subjects:
Cell Adhesion
Erythrocytes - cytology
Erythrocytes - immunology
Fc γ receptors
Flow Cytometry - methods
Fluorescent Dyes
Humans
Immunophenotyping
Organic Chemicals
Phagocytosis
PKH26
Receptors, Fc - immunology
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Summary:Human receptors for IgG (Fc γR) are characterized by diverse structure and function. We describe a flow cytometric technique to quantitate receptor-specific Fc γR-mediated attachment and phagocytosis in phenotypically-defined subsets of cells using erythrocytes (E) labeled with the lipophilic dye PKH26 and coupled with biotin/avidin to either human IgG (myeloma proteins) or anti-Fc γR mAb. Using this technique and Fc γRIIa as a model, (1) we demonstrate sensitive and specific quantitation of attached and internalized E coupled to receptor-specific mAb or natural ligand by monocytes within a peripheral blood leukocyte preparation; (2) we show the capacity to detect subtle allelic differences in Fc γR function; and (3) we demonstrate oxidant-induced enhancement of binding and internalization.
ISSN:0022-1759
1872-7905
DOI:10.1016/S0022-1759(97)00053-7