Macrophage-derived neutrophil chemotactic factor is involved in the neutrophil recruitment inhibitory activity present in the supernatants of LPS-stimulated macrophages
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...
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| Format: | Article |
| Language: | English |
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Wiley
1996-01-01
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| Series: | Mediators of Inflammation |
| Online Access: | http://dx.doi.org/10.1155/S0962935196000208 |
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| author | B. M. Tavares-Murta F. Q. Cunha M. Dias-Baruffi M. C. Roque-Barreira S. H. Ferreira |
| author_facet | B. M. Tavares-Murta F. Q. Cunha M. Dias-Baruffi M. C. Roque-Barreira S. H. Ferreira |
| author_sort | B. M. Tavares-Murta |
| collection | DOAJ |
| description | 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-α 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-α 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-α and IL-8, and that MNCF accounts for the inhibition of neutrophil migration in vivo by the D-gal+ fraction. |
| format | Article |
| id | doaj-art-d9d21bd40a9b452ca3e081ab0e812489 |
| institution | Kabale University |
| issn | 0962-9351 1466-1861 |
| language | English |
| publishDate | 1996-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Mediators of Inflammation |
| spelling | doaj-art-d9d21bd40a9b452ca3e081ab0e8124892025-02-03T01:13:14ZengWileyMediators of Inflammation0962-93511466-18611996-01-015211612010.1155/S0962935196000208Macrophage-derived neutrophil chemotactic factor is involved in the neutrophil recruitment inhibitory activity present in the supernatants of LPS-stimulated macrophagesB. M. Tavares-Murta0F. Q. Cunha1M. Dias-Baruffi2M. C. Roque-Barreira3S. H. Ferreira4Department of Pharmacology, Faculty of Medicine of Ribeirão Preto, USP, Ribeirão Preto, SP 14049-900, BrazilDepartment of Pharmacology, Faculty of Medicine of Ribeirão Preto, USP, Ribeirão Preto, SP 14049-900, BrazilDepartment of Parasitology, Microbiology and Immunology, Faculty of Medicine of Ribeirão Preto, USP, Ribeirão Preto, SP 14049-900, BrazilDepartment of Parasitology, Microbiology and Immunology, Faculty of Medicine of Ribeirão Preto, USP, Ribeirão Preto, SP 14049-900, BrazilDepartment of Pharmacology, Faculty of Medicine of Ribeirão Preto, USP, Ribeirão Preto, SP 14049-900, BrazilIn 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-α 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-α 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-α and IL-8, and that MNCF accounts for the inhibition of neutrophil migration in vivo by the D-gal+ fraction.http://dx.doi.org/10.1155/S0962935196000208 |
| spellingShingle | B. M. Tavares-Murta F. Q. Cunha M. Dias-Baruffi M. C. Roque-Barreira S. H. Ferreira Macrophage-derived neutrophil chemotactic factor is involved in the neutrophil recruitment inhibitory activity present in the supernatants of LPS-stimulated macrophages Mediators of Inflammation |
| title | Macrophage-derived neutrophil chemotactic factor is involved in the neutrophil recruitment inhibitory activity present in the supernatants of LPS-stimulated macrophages |
| title_full | Macrophage-derived neutrophil chemotactic factor is involved in the neutrophil recruitment inhibitory activity present in the supernatants of LPS-stimulated macrophages |
| title_fullStr | Macrophage-derived neutrophil chemotactic factor is involved in the neutrophil recruitment inhibitory activity present in the supernatants of LPS-stimulated macrophages |
| title_full_unstemmed | Macrophage-derived neutrophil chemotactic factor is involved in the neutrophil recruitment inhibitory activity present in the supernatants of LPS-stimulated macrophages |
| title_short | Macrophage-derived neutrophil chemotactic factor is involved in the neutrophil recruitment inhibitory activity present in the supernatants of LPS-stimulated macrophages |
| title_sort | macrophage derived neutrophil chemotactic factor is involved in the neutrophil recruitment inhibitory activity present in the supernatants of lps stimulated macrophages |
| url | http://dx.doi.org/10.1155/S0962935196000208 |
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