Isolation and functional analysis of neutrophils.

Robert A Clark, W. M. Nauseef

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

This unit describes the isolation of high-purity neutrophils (or polymorphonuclear leukocytes; PMN) and the assays that can be performed to assess their function. An assay for the in vitro study of phagocytic capacity is presented, and microbicidal assays using lysostaphin and differential centrifugation are also described. During phagocytosis or on exposure to soluble agonists, neutrophils exhibit a marked increase in oxidative metabolism which is mediated by activation of NADPH oxidase and results in the formation of superoxide anion as well as hydrogen peroxide and other reactive species. The nitroblue tetrazolium reduction slide test is presented, and is a nonquantitative screening test for the presence of reactive oxygen products. The quantitation of superoxide by the kinetic and static assays are detailed, and a "broken cell" assay is also outlined.Finally, measurement of hydrogen peroxide formation is described.

Original languageEnglish (US)
JournalCurrent protocols in immunology / edited by John E. Coligan ... [et al.]
VolumeChapter 7
StatePublished - May 2001
Externally publishedYes

Fingerprint

Neutrophils
Superoxides
Hydrogen Peroxide
Lysostaphin
Nitroblue Tetrazolium
NADPH Oxidase
Centrifugation
Phagocytosis
Oxygen
In Vitro Techniques

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Isolation and functional analysis of neutrophils. / Clark, Robert A; Nauseef, W. M.

In: Current protocols in immunology / edited by John E. Coligan ... [et al.], Vol. Chapter 7, 05.2001.

Research output: Contribution to journalArticle

@article{fd41931e5c3c4c3eacb6dfa10ad5d0d9,
title = "Isolation and functional analysis of neutrophils.",
abstract = "This unit describes the isolation of high-purity neutrophils (or polymorphonuclear leukocytes; PMN) and the assays that can be performed to assess their function. An assay for the in vitro study of phagocytic capacity is presented, and microbicidal assays using lysostaphin and differential centrifugation are also described. During phagocytosis or on exposure to soluble agonists, neutrophils exhibit a marked increase in oxidative metabolism which is mediated by activation of NADPH oxidase and results in the formation of superoxide anion as well as hydrogen peroxide and other reactive species. The nitroblue tetrazolium reduction slide test is presented, and is a nonquantitative screening test for the presence of reactive oxygen products. The quantitation of superoxide by the kinetic and static assays are detailed, and a {"}broken cell{"} assay is also outlined.Finally, measurement of hydrogen peroxide formation is described.",
author = "Clark, {Robert A} and Nauseef, {W. M.}",
year = "2001",
month = "5",
language = "English (US)",
volume = "Chapter 7",
journal = "Current Protocols in Immunology",
issn = "1934-3671",
publisher = "John Wiley and Sons Inc.",

}

TY - JOUR

T1 - Isolation and functional analysis of neutrophils.

AU - Clark, Robert A

AU - Nauseef, W. M.

PY - 2001/5

Y1 - 2001/5

N2 - This unit describes the isolation of high-purity neutrophils (or polymorphonuclear leukocytes; PMN) and the assays that can be performed to assess their function. An assay for the in vitro study of phagocytic capacity is presented, and microbicidal assays using lysostaphin and differential centrifugation are also described. During phagocytosis or on exposure to soluble agonists, neutrophils exhibit a marked increase in oxidative metabolism which is mediated by activation of NADPH oxidase and results in the formation of superoxide anion as well as hydrogen peroxide and other reactive species. The nitroblue tetrazolium reduction slide test is presented, and is a nonquantitative screening test for the presence of reactive oxygen products. The quantitation of superoxide by the kinetic and static assays are detailed, and a "broken cell" assay is also outlined.Finally, measurement of hydrogen peroxide formation is described.

AB - This unit describes the isolation of high-purity neutrophils (or polymorphonuclear leukocytes; PMN) and the assays that can be performed to assess their function. An assay for the in vitro study of phagocytic capacity is presented, and microbicidal assays using lysostaphin and differential centrifugation are also described. During phagocytosis or on exposure to soluble agonists, neutrophils exhibit a marked increase in oxidative metabolism which is mediated by activation of NADPH oxidase and results in the formation of superoxide anion as well as hydrogen peroxide and other reactive species. The nitroblue tetrazolium reduction slide test is presented, and is a nonquantitative screening test for the presence of reactive oxygen products. The quantitation of superoxide by the kinetic and static assays are detailed, and a "broken cell" assay is also outlined.Finally, measurement of hydrogen peroxide formation is described.

UR - http://www.scopus.com/inward/record.url?scp=52649112129&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=52649112129&partnerID=8YFLogxK

M3 - Article

VL - Chapter 7

JO - Current Protocols in Immunology

JF - Current Protocols in Immunology

SN - 1934-3671

ER -