A flow cytometric assay of Fc receptor-mediated phagocytosis

Wolfgang H.T. Schürmann; Mark W. Frampton; Gudrun Schürmann; Norbert J. Roberts; Joan E. Nichols; Jacob N. Finkelstein; Günter Oberdörster; Mark J. Utell

doi:10.1089/jam.1997.10.1

A flow cytometric assay of Fc receptor-mediated phagocytosis

Wolfgang H.T. Schürmann, Mark W. Frampton, Gudrun Schürmann, Norbert J. Roberts, Joan E. Nichols, Jacob N. Finkelstein, Günter Oberdörster, Mark J. Utell

Internal Medicine

Research output: Contribution to journal › Article › peer-review

4 Scopus citations

Abstract

A flow cytometric assay was developed that specifically measures Fc receptor-mediated phagocytosis independent of other phagocytosis-related receptors. Suspensions of rat alveolar macrophages were incubated with fluorescein isothiocyanate (FITC)labeled polystyrene microspheres and subsequently were analyzed by flow cytometry. Coating of microspheres with bovine serum albumin was used to generate 'neutral' particles demonstrating low phagocytosis by alveolar macrophages (AMs). AM attachment and uptake of these particles was increased tenfold with IgG opsonization. This increase was abolished completely by blocking of AM Fc receptors with IgG. The quenching effect of ethidium bromide (EB) on FITC fluorescence was used for differentiating attached from internalized particles. The assay was standardized and optimized for use in toxicologic studies.

Original language	English (US)
Pages (from-to)	1-8
Number of pages	8
Journal	Journal of Aerosol Medicine: Deposition, Clearance, and Effects in the Lung
Volume	10
Issue number	1
DOIs	https://doi.org/10.1089/jam.1997.10.1
State	Published - 1997

Keywords

Fc receptor
alveolar macrophages
flow cytometry
particles
phagocytosis

ASJC Scopus subject areas

Pulmonary and Respiratory Medicine
Pharmacology (medical)

Access to Document

10.1089/jam.1997.10.1

Cite this

@article{bf141169e851447bbbf9b5ffaf64573a,

title = "A flow cytometric assay of Fc receptor-mediated phagocytosis",

abstract = "A flow cytometric assay was developed that specifically measures Fc receptor-mediated phagocytosis independent of other phagocytosis-related receptors. Suspensions of rat alveolar macrophages were incubated with fluorescein isothiocyanate (FITC)labeled polystyrene microspheres and subsequently were analyzed by flow cytometry. Coating of microspheres with bovine serum albumin was used to generate 'neutral' particles demonstrating low phagocytosis by alveolar macrophages (AMs). AM attachment and uptake of these particles was increased tenfold with IgG opsonization. This increase was abolished completely by blocking of AM Fc receptors with IgG. The quenching effect of ethidium bromide (EB) on FITC fluorescence was used for differentiating attached from internalized particles. The assay was standardized and optimized for use in toxicologic studies.",

keywords = "Fc receptor, alveolar macrophages, flow cytometry, particles, phagocytosis",

author = "Sch{\"u}rmann, {Wolfgang H.T.} and Frampton, {Mark W.} and Gudrun Sch{\"u}rmann and Roberts, {Norbert J.} and Nichols, {Joan E.} and Finkelstein, {Jacob N.} and G{\"u}nter Oberd{\"o}rster and Utell, {Mark J.}",

year = "1997",

doi = "10.1089/jam.1997.10.1",

language = "English (US)",

volume = "10",

pages = "1--8",

journal = "Journal of Aerosol Medicine: Deposition, Clearance, and Effects in the Lung",

issn = "0894-2684",

publisher = "Mary Ann Liebert Inc.",

number = "1",

}

TY - JOUR

T1 - A flow cytometric assay of Fc receptor-mediated phagocytosis

AU - Schürmann, Wolfgang H.T.

AU - Frampton, Mark W.

AU - Schürmann, Gudrun

AU - Roberts, Norbert J.

AU - Nichols, Joan E.

AU - Finkelstein, Jacob N.

AU - Oberdörster, Günter

AU - Utell, Mark J.

PY - 1997

Y1 - 1997

N2 - A flow cytometric assay was developed that specifically measures Fc receptor-mediated phagocytosis independent of other phagocytosis-related receptors. Suspensions of rat alveolar macrophages were incubated with fluorescein isothiocyanate (FITC)labeled polystyrene microspheres and subsequently were analyzed by flow cytometry. Coating of microspheres with bovine serum albumin was used to generate 'neutral' particles demonstrating low phagocytosis by alveolar macrophages (AMs). AM attachment and uptake of these particles was increased tenfold with IgG opsonization. This increase was abolished completely by blocking of AM Fc receptors with IgG. The quenching effect of ethidium bromide (EB) on FITC fluorescence was used for differentiating attached from internalized particles. The assay was standardized and optimized for use in toxicologic studies.

AB - A flow cytometric assay was developed that specifically measures Fc receptor-mediated phagocytosis independent of other phagocytosis-related receptors. Suspensions of rat alveolar macrophages were incubated with fluorescein isothiocyanate (FITC)labeled polystyrene microspheres and subsequently were analyzed by flow cytometry. Coating of microspheres with bovine serum albumin was used to generate 'neutral' particles demonstrating low phagocytosis by alveolar macrophages (AMs). AM attachment and uptake of these particles was increased tenfold with IgG opsonization. This increase was abolished completely by blocking of AM Fc receptors with IgG. The quenching effect of ethidium bromide (EB) on FITC fluorescence was used for differentiating attached from internalized particles. The assay was standardized and optimized for use in toxicologic studies.

KW - Fc receptor

KW - alveolar macrophages

KW - flow cytometry

KW - particles

KW - phagocytosis

UR - http://www.scopus.com/inward/record.url?scp=0030889115&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030889115&partnerID=8YFLogxK

U2 - 10.1089/jam.1997.10.1

DO - 10.1089/jam.1997.10.1

M3 - Article

AN - SCOPUS:0030889115

SN - 0894-2684

VL - 10

SP - 1

EP - 8

JO - Journal of Aerosol Medicine: Deposition, Clearance, and Effects in the Lung

JF - Journal of Aerosol Medicine: Deposition, Clearance, and Effects in the Lung

IS - 1

ER -

A flow cytometric assay of Fc receptor-mediated phagocytosis

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this