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

Human receptors for IgG (Fc gamma R) are characterized by diverse structure and function. We describe a flow cytometric technique to quantitate receptor-specific Fc gamma R-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 gamma R mAb. Using this technique and Fc gamma RIIa 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 gamma R function; and (3) we demonstrate oxidant-induced enhancement of binding and internalization.