Isolation and partial chemical characterization of macrophage-derived neutrophil chemotactic factor

Macrophages stimulated with lipopolysaccharide (LPS) release a factor (MNCF; macrophage-derived neutrophil chemotactic factor) which induces neutrophil migration in vivo and in vitro. The in vivo chemotactic activity of crude MNCF is not affected by pretreating the animals with dexamethasone, an unc...

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Main Authors: M. Dias-Baruffi, M. C. Roque-Barreira, F. Q. Cunha, S. H. Ferreira
Format: Article
Language:English
Published: Wiley 1995-01-01
Series:Mediators of Inflammation
Online Access:http://dx.doi.org/10.1155/S096293519500041X
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author M. Dias-Baruffi
M. C. Roque-Barreira
F. Q. Cunha
S. H. Ferreira
author_facet M. Dias-Baruffi
M. C. Roque-Barreira
F. Q. Cunha
S. H. Ferreira
author_sort M. Dias-Baruffi
collection DOAJ
description Macrophages stimulated with lipopolysaccharide (LPS) release a factor (MNCF; macrophage-derived neutrophil chemotactic factor) which induces neutrophil migration in vivo and in vitro. The in vivo chemotactic activity of crude MNCF is not affected by pretreating the animals with dexamethasone, an uncommon characteristic which discriminates MNCF from known chemotactic cytokines. We purified MNCF by affinity chromatography of the supernatant from LPS-stimulated macrophages on immobilized D-galactose, followed by gel filtration of the sugar-binding material on Superdex 75. The activity was eluted in the volume corresponding to a MW of 54 kDa. SDS–PAGE of this preparation revealed a single band, also corresponding to a 54 kDa protein. MNCF is an acidic protein (pI < 4) as shown by chromatofocussing. Like the crude MNCF, the homogeneous protein induced neutrophil migration in vitro as well as in vivo. This was not modified by dexamethasone pretreatment.
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publishDate 1995-01-01
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series Mediators of Inflammation
spelling doaj-art-5a44a200ac8144db8b8635c9fa42c9ab2025-08-20T02:05:28ZengWileyMediators of Inflammation0962-93511466-18611995-01-014425726210.1155/S096293519500041XIsolation and partial chemical characterization of macrophage-derived neutrophil chemotactic factorM. Dias-Baruffi0M. C. Roque-Barreira1F. Q. Cunha2S. H. Ferreira3Department of Immunology, Faculty of Medicine, São Paulo, 14, Ribeirão Preto 049-900, BrazilDepartment of Immunology, Faculty of Medicine, São Paulo, 14, Ribeirão Preto 049-900, BrazilDepartment of pharmacology, Faculty of Medicine, São Paulo, 14, Ribeirão Preto 049-900, BrazilDepartment of pharmacology, Faculty of Medicine, São Paulo, 14, Ribeirão Preto 049-900, BrazilMacrophages stimulated with lipopolysaccharide (LPS) release a factor (MNCF; macrophage-derived neutrophil chemotactic factor) which induces neutrophil migration in vivo and in vitro. The in vivo chemotactic activity of crude MNCF is not affected by pretreating the animals with dexamethasone, an uncommon characteristic which discriminates MNCF from known chemotactic cytokines. We purified MNCF by affinity chromatography of the supernatant from LPS-stimulated macrophages on immobilized D-galactose, followed by gel filtration of the sugar-binding material on Superdex 75. The activity was eluted in the volume corresponding to a MW of 54 kDa. SDS–PAGE of this preparation revealed a single band, also corresponding to a 54 kDa protein. MNCF is an acidic protein (pI < 4) as shown by chromatofocussing. Like the crude MNCF, the homogeneous protein induced neutrophil migration in vitro as well as in vivo. This was not modified by dexamethasone pretreatment.http://dx.doi.org/10.1155/S096293519500041X
spellingShingle M. Dias-Baruffi
M. C. Roque-Barreira
F. Q. Cunha
S. H. Ferreira
Isolation and partial chemical characterization of macrophage-derived neutrophil chemotactic factor
Mediators of Inflammation
title Isolation and partial chemical characterization of macrophage-derived neutrophil chemotactic factor
title_full Isolation and partial chemical characterization of macrophage-derived neutrophil chemotactic factor
title_fullStr Isolation and partial chemical characterization of macrophage-derived neutrophil chemotactic factor
title_full_unstemmed Isolation and partial chemical characterization of macrophage-derived neutrophil chemotactic factor
title_short Isolation and partial chemical characterization of macrophage-derived neutrophil chemotactic factor
title_sort isolation and partial chemical characterization of macrophage derived neutrophil chemotactic factor
url http://dx.doi.org/10.1155/S096293519500041X
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AT shferreira isolationandpartialchemicalcharacterizationofmacrophagederivedneutrophilchemotacticfactor