Evaluation of intracellular killing of bacteria by enriched populations of mouse peritoneal exudate neutrophils

P. H. Hart, L. K. Spencer, P. J. McDonald, J. J. Finlay-Jones

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Elicited, mouse peritoneal exudate cells were fractionated by centrifugation on discontinuous Percoll density gradients. Two subpopulations of neutrophils, each of greater than 90% purity, were isolated at discontinuous density gradient interfaces different from the region of mononuclear cell enrichment (i.e. 1.0694-1.0871 and 1.0872-1.1002 g/ml for neutrophils and less than 1.0694 g/ml for mononuclear cells). Peritoneal exudate cells were mixed with Proteus mirabilis in the presence of 1% normal mouse serum for 30 min. The mixtures were fractionated on gradients of Percoll diluted with a calcium-free medium. Populations of cells banding at densities greater than 1.0693 g/ml were washed free of gradient material, and neutrophil suspensions containing intracellular bacteria and which were relatively free of extracellular bacteria were isolated. Less than 7% of the total bacteria present was extracellular. The continuing extracellular presence of a heat-labile component of normal mouse serum was essential for maximal intracellular kill of P. mirabilis by mouse peritoneal neutrophils.

Original languageEnglish
Pages (from-to)361-370
Number of pages10
JournalAustralian Journal of Experimental Biology and Medical Science
Volume63
Issue number4
DOIs
Publication statusPublished - 1 Jan 1985
Externally publishedYes

Fingerprint

Exudates and Transudates
Bacteria
Neutrophils
Proteus mirabilis
Population
Centrifugation
Suspensions
Cells
Calcium
Serum
Hot Temperature
Percoll

Cite this

@article{aa0e7de721424ab482933cb5723ccd69,
title = "Evaluation of intracellular killing of bacteria by enriched populations of mouse peritoneal exudate neutrophils",
abstract = "Elicited, mouse peritoneal exudate cells were fractionated by centrifugation on discontinuous Percoll density gradients. Two subpopulations of neutrophils, each of greater than 90{\%} purity, were isolated at discontinuous density gradient interfaces different from the region of mononuclear cell enrichment (i.e. 1.0694-1.0871 and 1.0872-1.1002 g/ml for neutrophils and less than 1.0694 g/ml for mononuclear cells). Peritoneal exudate cells were mixed with Proteus mirabilis in the presence of 1{\%} normal mouse serum for 30 min. The mixtures were fractionated on gradients of Percoll diluted with a calcium-free medium. Populations of cells banding at densities greater than 1.0693 g/ml were washed free of gradient material, and neutrophil suspensions containing intracellular bacteria and which were relatively free of extracellular bacteria were isolated. Less than 7{\%} of the total bacteria present was extracellular. The continuing extracellular presence of a heat-labile component of normal mouse serum was essential for maximal intracellular kill of P. mirabilis by mouse peritoneal neutrophils.",
author = "Hart, {P. H.} and Spencer, {L. K.} and McDonald, {P. J.} and Finlay-Jones, {J. J.}",
year = "1985",
month = "1",
day = "1",
doi = "10.1038/icb.1985.42",
language = "English",
volume = "63",
pages = "361--370",
journal = "Immunology & Cell Biology",
issn = "0818-9641",
publisher = "Nature Publishing Group - Macmillan Publishers",
number = "4",

}

Evaluation of intracellular killing of bacteria by enriched populations of mouse peritoneal exudate neutrophils. / Hart, P. H.; Spencer, L. K.; McDonald, P. J.; Finlay-Jones, J. J.

In: Australian Journal of Experimental Biology and Medical Science, Vol. 63, No. 4, 01.01.1985, p. 361-370.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Evaluation of intracellular killing of bacteria by enriched populations of mouse peritoneal exudate neutrophils

AU - Hart, P. H.

AU - Spencer, L. K.

AU - McDonald, P. J.

AU - Finlay-Jones, J. J.

PY - 1985/1/1

Y1 - 1985/1/1

N2 - Elicited, mouse peritoneal exudate cells were fractionated by centrifugation on discontinuous Percoll density gradients. Two subpopulations of neutrophils, each of greater than 90% purity, were isolated at discontinuous density gradient interfaces different from the region of mononuclear cell enrichment (i.e. 1.0694-1.0871 and 1.0872-1.1002 g/ml for neutrophils and less than 1.0694 g/ml for mononuclear cells). Peritoneal exudate cells were mixed with Proteus mirabilis in the presence of 1% normal mouse serum for 30 min. The mixtures were fractionated on gradients of Percoll diluted with a calcium-free medium. Populations of cells banding at densities greater than 1.0693 g/ml were washed free of gradient material, and neutrophil suspensions containing intracellular bacteria and which were relatively free of extracellular bacteria were isolated. Less than 7% of the total bacteria present was extracellular. The continuing extracellular presence of a heat-labile component of normal mouse serum was essential for maximal intracellular kill of P. mirabilis by mouse peritoneal neutrophils.

AB - Elicited, mouse peritoneal exudate cells were fractionated by centrifugation on discontinuous Percoll density gradients. Two subpopulations of neutrophils, each of greater than 90% purity, were isolated at discontinuous density gradient interfaces different from the region of mononuclear cell enrichment (i.e. 1.0694-1.0871 and 1.0872-1.1002 g/ml for neutrophils and less than 1.0694 g/ml for mononuclear cells). Peritoneal exudate cells were mixed with Proteus mirabilis in the presence of 1% normal mouse serum for 30 min. The mixtures were fractionated on gradients of Percoll diluted with a calcium-free medium. Populations of cells banding at densities greater than 1.0693 g/ml were washed free of gradient material, and neutrophil suspensions containing intracellular bacteria and which were relatively free of extracellular bacteria were isolated. Less than 7% of the total bacteria present was extracellular. The continuing extracellular presence of a heat-labile component of normal mouse serum was essential for maximal intracellular kill of P. mirabilis by mouse peritoneal neutrophils.

UR - http://www.scopus.com/inward/record.url?scp=0022412108&partnerID=8YFLogxK

U2 - 10.1038/icb.1985.42

DO - 10.1038/icb.1985.42

M3 - Article

VL - 63

SP - 361

EP - 370

JO - Immunology & Cell Biology

JF - Immunology & Cell Biology

SN - 0818-9641

IS - 4

ER -