scholarly journals Biological characterization of purified macrophage-derived neutrophil chemotactic factor

1995 ◽  
Vol 4 (4) ◽  
pp. 263-269 ◽  
Author(s):  
M. Dias-Baruffi ◽  
M. C. Roque-Barreira ◽  
F. Q. Cunha ◽  
S. H. Ferreira
Keyword(s):  
Neutrophil Migration ◽  
Chemotactic Activity ◽  
Chemotactic Factor ◽  
Biological Characterization ◽  
Important Mediator ◽  
Neutrophil Chemotactic Factor ◽  
Chemotactic Gradient

We have recently described the purification of a 54 kDa acidic protein, identified as macrophage-derived neutrophil chemotactic factor (MNCF). This protein causesin vitrochemotaxis as well asin vivoneutrophil migration even in animals treated with dexamethasone. Thisin vivochemotactic activity of MNCF in animals pretreated with dexamethasone is an uncommon characteristic which discriminates MNCF from known chemotactic cytokines. MNCF is released in the supernatant by macrophage monolayers stimulated with lipopolysaccharide (LPS). In the present study, we describe some biological characteristics of homogenous purified MNCF. When assayedin vitro, MNCF gave a bell-shaped dose–response curve. Thisin vitroactivity was shown to be caused by haptotaxis. Unlike N-formyl-methionylleucyl- phenylalanine (FMLP) or interleukin 8 (IL-8), the chemotactic activity of MNCFin vivoandin vitro, was inhibited by preincubation with D-galactose but not with D-mannose. In contrast with IL-8, MNCF did not bind to heparin and antiserum against IL-8 was ineffective in inhibiting its chemotactic activity. These data indicate that MNCF induces neutrophil migration through a carbohydrate recognition property, but by a mechanism different from that of the known chemokines. It is suggested that MNCF may be an important mediator in the recruitment of neutrophils via the formation of a substrate bound chemotactic gradient (haptotaxis) in the inflamed tissues.

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

1995 ◽  
Vol 4 (4) ◽  
pp. 257-262 ◽  
Author(s):  
M. Dias-Baruffi ◽  
M. C. Roque-Barreira ◽  
F. Q. Cunha ◽  
S. H. Ferreira
Keyword(s):  
Neutrophil Migration ◽  
Gel Filtration ◽  
Chemical Characterization ◽  
Chemotactic Factor ◽  
Sds Page ◽  
Neutrophil Chemotactic Factor ◽  
Partial Chemical

Macrophages stimulated with lipopolysaccharide (LPS) release a factor (MNCF; macrophage-derived neutrophil chemotactic factor) which induces neutrophil migrationin vivoandin vitro. Thein vivochemotactic 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 migrationin vitroas well asin vivo. This was not modified by dexamethasone pretreatment.

scholarly journals The Macrophage-derived Lectin, MNCF, Activates Neutrophil Migration through a Pertussis Toxin-sensitive Pathway

2005 ◽  
Vol 53 (6) ◽  
pp. 715-723 ◽  
Author(s):  
Andréa N. Moreno ◽  
Gabriela Pereira-da-Silva ◽  
Constance Oliver ◽  
Maria Célia Jamur ◽  
Ademilson Panunto-Castelo ◽  
...  
Keyword(s):  
Cell Surface ◽  
Pertussis Toxin ◽  
Neutrophil Migration ◽  
Cell Polarization ◽  
Glucocorticoid Treatment ◽  
Neutrophil Chemotactic Factor ◽  
G Protein Coupled ◽  
Neutrophil Surface

The macrophage-derived neutrophil chemotactic factor (MNCF) is a d-galactose-binding lectin that induces neutrophil migration in vitro and in vivo. Neutrophil recruitment induced by MNCF is resistant to glucocorticoid treatment and is inhibited by the lectin-specific sugar, d-galactose. In the present study, we characterized the binding of MNCF to neutrophils and the responses triggered by this binding. Exposure to MNCF resulted in cell polarization, formation of a lamellipodium, and deep ruffles on the cell surface. By confocal microscopy, we observed that MNCF was evenly distributed on the cell surface after 30 min of incubation. The labeling intensity progressively diminished with longer incubations. Internalization kinetics showed that MNCF/ligand complexes were rapidly internalized, reaching maximum intracellular concentrations at 120 min and then decreased thereafter. The binding and internalization of MNCF were selectively inhibited by d-galactose. MNCF-induced neutrophil chemotaxis was inhibited by pertussis toxin. This fact strongly suggests that the MNCF-ligand on the neutrophil surface is a G-protein-coupled receptor (GPCR), similar to receptors for well-established neutrophil attractants. Our observations on the ability of MNCF to activate neutrophils are consistent with the increasing evidence for the participation of animal lectins in the innate immune response.

In vitro and in vivo biological characterization of the anti-proliferative potential of a cyclic trinuclear organotin(iv) complex

2016 ◽  
Vol 12 (3) ◽  
pp. 1015-1023 ◽  
Author(s):  
Marta Martins ◽  
Pedro V. Baptista ◽  
Ana Soraia Mendo ◽  
Claudia Correia ◽  
Paula Videira ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Proliferative Potential ◽  
Biological Characterization ◽  
Growth Of Tumor

Identification of novel molecules that can selectively inhibit the growth of tumor cells, is of utmost importance.

scholarly journals The Biology of COP Cells: Mesenchymal of Hematopoietic?

2021 ◽  
Vol 5 (Supplement_1) ◽  
pp. 45-45
Author(s):  
Meghan McGee-Lawrence
Keyword(s):  
Stem Cells ◽  
Clinical Practice ◽  
Progenitor Cells ◽  
Peripheral Blood ◽  
Biological Characterization ◽  
Differentiation Capacity ◽  
Growing Body

Abstract Circulating osteogenic precursor (COP) cells constitute a recently discovered population of circulating progenitor cells with the capacity to form not only bone but other mesenchymal tissues. A small but growing body of literature explores these cells, but with a great deal of disagreement and contradiction within it, mainly whether these cells are from mesenchymal or hematopoietic origin. This session will discuss the origins and biological characterization of these cells, including the identification strategies used to isolate these cells from the peripheral blood. It also examines the available knowledge on the in vitro and in vivo behaviour of these cells in plastic adherence, differentiation capacity, proliferation, and cellular homing. We will also review the profound and exciting implications for future use of COP cells in clinical practice, particularly in comparison with other types of stem cells.

Chicken mim-1 Protein, P33, Is a Heterophil Chemotactic Factor Present in Salmonella Enteritidis Immune Lymphokine

2001 ◽  
Vol 64 (10) ◽  
pp. 1503-1509 ◽  
Author(s):  
K. M. BISCHOFF ◽  
E. J. PISHKO ◽  
K. J. GENOVESE ◽  
T. L. CRIPPEN ◽  
C. K. HOLTZAPPLE ◽  
...  
Keyword(s):  
Salmonella Enteritidis ◽  
Polyclonal Antibodies ◽  
Active Role ◽  
Repeat Sequence ◽  
Chemotactic Factor ◽  
Acid Protein ◽  
Neutrophil Chemotactic Factor ◽  
Amino Acid Protein

Lymphokine (ILK) secreted from concanavalin A-stimulated T cells from Salmonella Enteritidis-immune chickens is an undefined mixture of proteins that confers protection against Salmonella infectivity when administered to day-old chicks. It has previously been shown that polyclonal antibodies raised against human granulocyte colony-stimulating factor (GCSF) can neutralize the heterophil activation that is responsible for ILK's protective effect. Western blot analysis of ILK probed with anti-GCSF antibodies detects a prominent protein of mass 33 kDa. We have sequenced the first 20 amino acids of this protein and found it to be identical to residues 24 to 43 of P33, a 326-amino acid protein of unknown function encoded by the chicken mim-1 gene. The primary structure of P33 consists of two 140-residue imperfect repeats that are each homologous to a mammalian neutrophil chemotactic factor termed leukocyte cell-derived chemotaxin 2 (LECT2). We have expressed mim-1 in Escherichia coli and demonstrated in vitro that recombinant P33 is chemotactic for heterophils, the avian equivalent of mammalian neutrophils. We have also constructed a derivative of P33 that consists of residues 33 to 165 (P33[33–165]), the first repeat sequence of P33 that is homologous to LECT2. P33(33–165) is chemotactic for heterophils both in vitro and in vivo, inducing an influx of heterophils into the peritoneum in a response similar to that observed with ILK. These results suggest that P33 functions as a chemotactic factor in chickens and that it plays an active role in ILK-mediated protection against Salmonella infection.

scholarly journals Macrophage-derived neutrophil chemotactic factor is involved in the neutrophil recruitment inhibitory activity present in the supernatants of LPS-stimulated macrophages

1996 ◽  
Vol 5 (2) ◽  
pp. 116-120 ◽  
Author(s):  
B. M. Tavares-Murta ◽  
F. Q. Cunha ◽  
M. Dias-Baruffi ◽  
M. C. Roque-Barreira ◽  
S. H. Ferreira
Keyword(s):  
Affinity Chromatography ◽  
Inhibitory Activity ◽  
Neutrophil Migration ◽  
Gel Filtration ◽  
Inhibitory Effect ◽  
Neutrophil Recruitment ◽  
Chemotactic Factor ◽  
Tnf Α ◽  
Neutrophil Chemotactic Factor

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 migrationin vivoby the D-gal+fraction.

scholarly journals Biological characterization of 2-aminothiazole-derived Cdk4/6 selective inhibitor in vitro and in vivo

Cell Cycle ◽  
2010 ◽  
Vol 9 (8) ◽  
pp. 1590-1600 ◽  
Author(s):  
Hiroshi Hirai ◽  
Toshiyasu Shimomura ◽  
Makiko Kobayashi ◽  
Tomohiro Eguchi ◽  
Eri Taniguchi ◽  
...  
Keyword(s):  
Selective Inhibitor ◽  
Biological Characterization

Biological Characterization of Ingredients in OPLC-BioArena-Greenhouse-System: Unique Reactions of Endogenous HCHO and O3 in In Vitro and In Vivo Conditions

2012 ◽  
Vol 75 (17) ◽  
pp. 983-990 ◽  
Author(s):  
Ernő Tyihák ◽  
Ágnes M. Móricz ◽  
Péter G. Ott ◽  
György Kátay ◽  
Emil Mincsovics
Keyword(s):  
Biological Characterization

Interleukin-8 is a major component of pleural liquid chemotactic activity in a rabbit model of endotoxin pleurisy

1994 ◽  
Vol 267 (2) ◽  
pp. L137-L144 ◽  
Author(s):  
A. M. Boylan ◽  
C. A. Hebert ◽  
M. Sadick ◽  
W. L. Wong ◽  
A. Chuntharapai ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Monoclonal Antibodies ◽  
Neutrophil Migration ◽  
Rabbit Model ◽  
Pleural Space ◽  
Chemotactic Activity ◽  
Interleukin 8 ◽  
Gram Negative

Gram-negative endotoxin induces production of the potent chemotactic factor interleukin-8 (IL-8) in vitro; however, the importance of IL-8 in endotoxin-induced inflammation in vivo is unknown. We asked whether IL-8 is an important contributor to chemotactic activity in acute inflammatory liquids formed in response to endotoxin, and, if present, what concentrations of IL-8 antigen are generated. For these studies, we cloned and expressed rabbit recombinant IL-8 (rrIL-8), developed specific anti-rabbit IL-8 monoclonal antibodies (mAb), and then used these reagents to develop assays to detect rabbit IL-8 bioactivity and measure rabbit IL-8 antigen. Escherichia coli endotoxin (20 ng/ml, n = 4, or 2,000 ng/ml, n = 4) was instilled into the pleural space of eight rabbits for 6 h. Rabbit IL-8 bioactivity in the endotoxin pleurisy samples was assayed by measuring the migration of rabbit neutrophils toward the pleural liquid under two different conditions: 1) after addition of an anti-IL-8 neutralizing mAb and 2) after desensitization of the neutrophils to rrIL-8. Addition of the anti-IL-8 mAb decreased neutrophil migration toward the pleural liquid by 65 +/- 13 and 75 +/- 22% (mean +/- SE, after 20 and 2,000 ng/ml endotoxin, respectively; P < 0.01 compared with a control mAb). Desensitization of neutrophils to rrIL-8 decreased their migration toward the pleural liquid by 72 +/- 5% (P = 0.03, compared with exposure of neutrophils to buffer alone.(ABSTRACT TRUNCATED AT 250 WORDS)

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