Flow cytometric detection and quantitative analysis of the GM-CSF receptor in human granulocytes and comparison with the radioligand binding assay

Alessandra Stacchini, Lidia Fubini, Massimo Aglietta

Research output: Contribution to journalArticle

Abstract

A flow cytometric method to quantify the granulocyte-macrophage colony-stimulating factor receptor (GM-CSFr) on human cells is described. The number of GM-CSFr binding sites on human neutrophils was assessed by using different bead standards. Results were compared with those from conventional receptor quantification, which was performed by using the radioligand binding assay. A high degree of correlation was found between the two methods, although quantitative evaluation of GM-CSFr expression on neutrophils performed by flow cytometry revealed a somewhat higher number of receptor molecules per cell than with that determined by Scatchard analysis. By the flow quantitative approach, we measured the GM-CSFr on mobilized CD34-positive cells and obtained results similar to those of previously published data. Our data suggest that flow cytometric analysis is a simple and reproducible method to detect and quantify the presence of GM-CSFr per cell, thus allowing the study of receptor expression on different populations selected by gating on the basis of the scatter parameters and surface markers. This assay offers the possibility to quantify the presence of GM-CSFr on different subsets of normal and pathological cells even if samples are too small (such as CD34-positive progenitor cells) for measurement with the radioligand binding assay.

Original languageEnglish
Pages (from-to)374-381
Number of pages8
JournalCytometry
Volume24
Issue number4
DOIs
Publication statusPublished - Aug 1 1996

Fingerprint

Granulocyte-Macrophage Colony-Stimulating Factor Receptors
Radioligand Assay
Granulocytes
Neutrophils
Flow Cytometry
Stem Cells
Binding Sites
Population

Keywords

  • GM-CSFr quantification
  • Microbeads
  • Radioligand binding

ASJC Scopus subject areas

  • Biophysics
  • Cell Biology
  • Endocrinology
  • Hematology
  • Pathology and Forensic Medicine

Cite this

Flow cytometric detection and quantitative analysis of the GM-CSF receptor in human granulocytes and comparison with the radioligand binding assay. / Stacchini, Alessandra; Fubini, Lidia; Aglietta, Massimo.

In: Cytometry, Vol. 24, No. 4, 01.08.1996, p. 374-381.

Research output: Contribution to journalArticle

@article{a926e6ddce104f4c99600eaab3c2a24a,
title = "Flow cytometric detection and quantitative analysis of the GM-CSF receptor in human granulocytes and comparison with the radioligand binding assay",
abstract = "A flow cytometric method to quantify the granulocyte-macrophage colony-stimulating factor receptor (GM-CSFr) on human cells is described. The number of GM-CSFr binding sites on human neutrophils was assessed by using different bead standards. Results were compared with those from conventional receptor quantification, which was performed by using the radioligand binding assay. A high degree of correlation was found between the two methods, although quantitative evaluation of GM-CSFr expression on neutrophils performed by flow cytometry revealed a somewhat higher number of receptor molecules per cell than with that determined by Scatchard analysis. By the flow quantitative approach, we measured the GM-CSFr on mobilized CD34-positive cells and obtained results similar to those of previously published data. Our data suggest that flow cytometric analysis is a simple and reproducible method to detect and quantify the presence of GM-CSFr per cell, thus allowing the study of receptor expression on different populations selected by gating on the basis of the scatter parameters and surface markers. This assay offers the possibility to quantify the presence of GM-CSFr on different subsets of normal and pathological cells even if samples are too small (such as CD34-positive progenitor cells) for measurement with the radioligand binding assay.",
keywords = "GM-CSFr quantification, Microbeads, Radioligand binding",
author = "Alessandra Stacchini and Lidia Fubini and Massimo Aglietta",
year = "1996",
month = "8",
day = "1",
doi = "10.1002/(SICI)1097-0320(19960801)24:4<374::AID-CYTO9>3.0.CO;2-F",
language = "English",
volume = "24",
pages = "374--381",
journal = "Cytometry Part B - Clinical Cytometry",
issn = "1552-4949",
publisher = "Wiley-Liss Inc.",
number = "4",

}

TY - JOUR

T1 - Flow cytometric detection and quantitative analysis of the GM-CSF receptor in human granulocytes and comparison with the radioligand binding assay

AU - Stacchini, Alessandra

AU - Fubini, Lidia

AU - Aglietta, Massimo

PY - 1996/8/1

Y1 - 1996/8/1

N2 - A flow cytometric method to quantify the granulocyte-macrophage colony-stimulating factor receptor (GM-CSFr) on human cells is described. The number of GM-CSFr binding sites on human neutrophils was assessed by using different bead standards. Results were compared with those from conventional receptor quantification, which was performed by using the radioligand binding assay. A high degree of correlation was found between the two methods, although quantitative evaluation of GM-CSFr expression on neutrophils performed by flow cytometry revealed a somewhat higher number of receptor molecules per cell than with that determined by Scatchard analysis. By the flow quantitative approach, we measured the GM-CSFr on mobilized CD34-positive cells and obtained results similar to those of previously published data. Our data suggest that flow cytometric analysis is a simple and reproducible method to detect and quantify the presence of GM-CSFr per cell, thus allowing the study of receptor expression on different populations selected by gating on the basis of the scatter parameters and surface markers. This assay offers the possibility to quantify the presence of GM-CSFr on different subsets of normal and pathological cells even if samples are too small (such as CD34-positive progenitor cells) for measurement with the radioligand binding assay.

AB - A flow cytometric method to quantify the granulocyte-macrophage colony-stimulating factor receptor (GM-CSFr) on human cells is described. The number of GM-CSFr binding sites on human neutrophils was assessed by using different bead standards. Results were compared with those from conventional receptor quantification, which was performed by using the radioligand binding assay. A high degree of correlation was found between the two methods, although quantitative evaluation of GM-CSFr expression on neutrophils performed by flow cytometry revealed a somewhat higher number of receptor molecules per cell than with that determined by Scatchard analysis. By the flow quantitative approach, we measured the GM-CSFr on mobilized CD34-positive cells and obtained results similar to those of previously published data. Our data suggest that flow cytometric analysis is a simple and reproducible method to detect and quantify the presence of GM-CSFr per cell, thus allowing the study of receptor expression on different populations selected by gating on the basis of the scatter parameters and surface markers. This assay offers the possibility to quantify the presence of GM-CSFr on different subsets of normal and pathological cells even if samples are too small (such as CD34-positive progenitor cells) for measurement with the radioligand binding assay.

KW - GM-CSFr quantification

KW - Microbeads

KW - Radioligand binding

UR - http://www.scopus.com/inward/record.url?scp=0029819190&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029819190&partnerID=8YFLogxK

U2 - 10.1002/(SICI)1097-0320(19960801)24:4<374::AID-CYTO9>3.0.CO;2-F

DO - 10.1002/(SICI)1097-0320(19960801)24:4<374::AID-CYTO9>3.0.CO;2-F

M3 - Article

C2 - 8866222

AN - SCOPUS:0029819190

VL - 24

SP - 374

EP - 381

JO - Cytometry Part B - Clinical Cytometry

JF - Cytometry Part B - Clinical Cytometry

SN - 1552-4949

IS - 4

ER -