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

Academic Article

Abstract

  • Human receptors for IgG (FcγR) are characterized by diverse structure and function. We describe a flow cytometric technique to quantitate receptor- specific FcγR-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 1gG (myeloma proteins) or anti-FcγR mAb. Using this technique and FcγRIIa 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 FcγR function; and (3) we demonstrate oxidant induced enhancement of binding and internalization.
  • Published In

    Digital Object Identifier (doi)

    Author List

  • Pricop L; Salmon JE; Edberg JC; Beavis AJ
  • Start Page

  • 55
  • End Page

  • 65
  • Volume

  • 205
  • Issue

  • 1