title: Macrophage-derived neutrophil chemotactic factor is involved in the neutrophil recruitment inhibitory activity present in the supernatants of LPS-stimulated macrophages.
abstract: In a previous study, we demonstrated the presence of a neutrophil recruitment inhibitory factor (NRIF) in the supernatants of LPS-stimulated macrophages. Recently, the purification of a 54 kDa protein, identified as the macrophage-derived neutrophil chemotactic factor (MNCF) was reported. Since NRIF and MNCF are obtained under the same conditions, and, since the intravenous administration of TNF-alpha and IL-8 inhibits neutrophil migration, we have investigated whether MNCF could be responsible for this inhibitory activity. After affinity chromatography of the macrophage supernatants on a D-galactose column, the inhibitory activity was recovered in both the unbound (D-gal(-)) and bound (D-gal(+)) fractions, with MNCF being found in the D-gal(+) fraction. Further gel filtration of the latter on Superdex 75 yielded a single peak containing both activities. In a cytotoxicity assay, most of the TNF found in the crude supernatants was recovered in the D-gal(-) fraction. Furthermore, the incubation of the D-gal(-) fraction with anti-TNF-alpha plus anti-IL-8 antisera partially prevents its inhibitory effect on neutrophil migration, but had no effect on the D-gal(+) activity. Overall, these results suggest that the D-gal(-) inhibitory effect is partially mediated by TNF-alpha and IL-8, and that MNCF accounts for the inhibition of neutrophil migration in vivo by the D-gal(+) fraction.
authors.affiliation: Department of Pharmacology Faculty of Medicine of Ribeirão Preto USP Ribeirão Preto SP 14049-900 Brazil.
authors.forename: B M
authors.initials: BM
authors.lastname: Tavares-Murta
authors.forename: F Q
authors.initials: FQ
authors.lastname: Cunha
authors.forename: M
authors.initials: M
authors.lastname: Dias-Baruffi
authors.forename: M C
authors.initials: MC
authors.lastname: Roque-Barreira
authors.forename: S H
authors.initials: SH
authors.lastname: Ferreira