Structure and Properties of a Novel Neutrophil-Activating Factor (NAF) Produced by Human Monocytes

Walz, Alfred; Peveri, Paola; Aschauer, H; Dewald, Béatrice; Baggiolini, Marco

doi:10.1007/978-3-642-73468-7_21

Cited by 3 publications

(4 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…All forms of NAF have four cysteine residues expected to form intrachain disulfide bridges, which appear important for activity because mercaptoethanol was found to be inhibitory (8). The disulfide bridges probably link the positions 7-34 and 9-50 as in 3-thromboglobulin (25) and platelet factor 4 (26), which are partially homologous to NAF (1,2,24).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Synthesis and expression in Escherichia coli of the gene encoding monocyte-derived neutrophil-activating factor: biological equivalence between natural and recombinant neutrophil-activating factor.

Lindley

Aschauer

Seifert

et al. 1988

Proc. Natl. Acad. Sci. U.S.A.

212

View full text Add to dashboard Cite

The neutrophil-activating factor (NAF) purified from the conditioned medium of lipopolysaccharidestimulated human monocytes was sequenced and found to consist of 72 amino acids: SAKELRCQCIKTYSKPFHPKFI-KELRVIESGPHCANTEIIVKLSDGRELCLDP-KENWVQRVVEKFLKRAENS. Purified preparations of natural NAF contained, in addition to this main form, minor amounts of three amino-terminal variants with 77 (+AVLPR), 70, and 69 residues. A gene coding for the 72-amino acid NAF was synthesized, cloned, and expressed in Escherichia coli. Western (immunologic) blot analysis of crude bacterial extracts, with an antiserum raised against natural NAF, revealed a single band that comigrated with natural NAF. Recombinant NAF purified to homogeneity had identical amino-and carboxyl-terminal sequences to the 72-amino acid natural NAF. Recombinant NAF was tested on human neutrophils and had the same activity and potency as natural NAF in inducing chemotaxis, rapidly increasing cytosolic free Ca2+, activating the respiratory burst, and releasing specific and azurophilic granular contents.

show abstract

Section: Discussionmentioning

confidence: 99%

“…Neutrophil-activating factor (NAF) is a peptide produced by human monocytes, which was purified recently (1)(2)(3). Independent work in different laboratories has established that peptides of this type are chemotactic for human neutrophils (4)(5)(6).…”

mentioning

confidence: 99%

Synthesis and expression in Escherichia coli of the gene encoding monocyte-derived neutrophil-activating factor: biological equivalence between natural and recombinant neutrophil-activating factor.

Lindley

Aschauer

Seifert

et al. 1988

Proc. Natl. Acad. Sci. U.S.A.

212

View full text Add to dashboard Cite

show abstract

“…These include potent chemokinetic and chemotactic activity (14-17), induction ofaggregation ( 15), and stimulation ofneutrophil lysosomal enzyme release (18). Another human AM product with a profound ability to modulate neutrophil function is a recently isolated (19,20), sequenced (21)(22)(23), and cloned (24) 72-residue neutrophil attractant/activation protein (NAP-1), which is also referred to as NAP-1/IL-8.2 Both NAP-I that is generated from lipopolysaccharide (LPS)-stimulated human peripheral blood monocytes (19) and recombinant NAP-I are potent chemotaxins for human neutrophils (25).…”

Section: Introductionmentioning

confidence: 99%

Macrophages cultured in vitro release leukotriene B4 and neutrophil attractant/activation protein (interleukin 8) sequentially in response to stimulation with lipopolysaccharide and zymosan.

Rankin¹,

Sylvester

Smith³

et al. 1990

J. Clin. Invest.

113

View full text Add to dashboard Cite

The capacity of lipopolysaccharide (LPS), zymosan, and calcium ionophore A23187 to induce neutrophil chemotactic activity (NCA), leukotriene B4 (LTB4), and neutrophil attractant/activation protein (NAP-1) release from human alveolar macrophages (AM) retrieved from normal nonsmokers was evaluated. LPS induced a dose-dependent release of LTB4 that began by 1 h, 4.0±3.2 ng/106 viable AM; peaked at 3 h, 24.7±13.5 ng/10' viable AM; and decreased by 24 h, 1.2±1.0 ng/106 viable AM (n = 8). Quantities of LTB4 in cell-free supernatants of AM stimulated with LPS were determined by reverse-phase high-performance liquid chromatography and corresponded well with results obtained by radioimmunoassay. By contrast, NAP-I release began -3-5 h after stimulation of AM with LPS, 197±192 ng/ml, and peaked at 24 h, 790±124 ng/ml. Release of NAP-1 was stimulus specific because A23187 evoked the release of LTB4 but not NAP-1, whereas LPS and zymosan induced the release of both LTB4 and NAP-1. The appearance of neutrophil chemotactic activity in supernatants of AM challenged with LPS for 3 h could be explained completely by the quantities of LTB4 present. After

show abstract

“…Paxillin phosphorylation in human neutrophils (A), CXCR1-RBL cells (B) and CXCR2-RBL cells (B, C) was induced by CXCL8 in concentrations that were shown in dose-dependent analyses to potently induce the migration of these cell types[19,20,[59][60][61]. (A) Neutrophils were cultured for 2 h on dishes coated by fibrinogen, with or without 100 ng/ml CXCL8, followed by paxillin immunoprecipitation.…”

mentioning

confidence: 99%

Cell migration to the chemokine CXCL8: Paxillin is activated and regulates adhesion and cell motility

Cohen-Hillel

Mintz

Meshel

et al. 2009

Cell. Mol. Life Sci.

View full text Add to dashboard Cite

The chemokine CXCL8 is a powerful inducer of directional cell motility, primarily during inflammation. In this study, we found that CXCL8 stimulation led to paxillin phosphorylation in normal neutrophils, and that both CXCL8 receptors (CXCR1 and CXCR2) mediated CXCL8-induced paxillin phosphorylation. In CXCR2-transfected cells, the process depended on G(alphai) and G(alphas) coupling to CXCR2. Dominant negative (DN) paxillin increased CXCL8-induced adhesion and migration, indicating that endogenous paxillin keeps migration at submaximal levels. Furthermore, using activating antibodies to beta1 integrins, analyses with focal adhesion kinase (FAK) DN variant (FRNK) and co-immunoprecipitations of FAK and paxillin, we found that beta1 integrin ligation cooperates with CXCL8-induced stimulation, leading to FAK activation and thereafter to FAK-mediated paxillin phosphorylation. Our findings indicate that paxillin keeps directional motility at a restrained magnitude, and suggest that perturbations in its activation may lead to chemotactic imbalance and to pathological conditions associated with excessive or reduced leukocyte migration.

show abstract

Structure and Properties of a Novel Neutrophil-Activating Factor (NAF) Produced by Human Monocytes

Cited by 3 publications

References 16 publications

Synthesis and expression in Escherichia coli of the gene encoding monocyte-derived neutrophil-activating factor: biological equivalence between natural and recombinant neutrophil-activating factor.

Synthesis and expression in Escherichia coli of the gene encoding monocyte-derived neutrophil-activating factor: biological equivalence between natural and recombinant neutrophil-activating factor.

Macrophages cultured in vitro release leukotriene B4 and neutrophil attractant/activation protein (interleukin 8) sequentially in response to stimulation with lipopolysaccharide and zymosan.

Cell migration to the chemokine CXCL8: Paxillin is activated and regulates adhesion and cell motility

Contact Info

Product

Resources

About