Production and Characterization of Recombinant Human Neutrophil Chemotactic Factor

Furuta, Ryuji; Yamagishi, Jun‐ichi; Kotani, Hirotada; Sakamoto, Fumiko; Fukui, Toshikazu; Y, Matsui; Sohmura, Yasunobu; Yamada, Masaaki; Yoshimura, Teizo; Larsen, Christian Grónhøj; Oppenheim, Joost J.; Matsushima, Kouji

doi:10.1093/oxfordjournals.jbchem.a122870

Cited by 63 publications

(24 citation statements)

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“…Moreover, human IL-8 was used in mouse inflammatory models with a neutrophil influx peaking at 3 hours. 45 Our data indicate that short-term exposure to fever-like temperatures did not affect adhesion and spreading of GM-CSF-and IL-8-treated neutrophils to fibronectin but inhibited neutrophil migration. In accordance with our data, Frohlich and colleagues 46 demonstrated that heat did not prevent neutrophil adhesion to IL-1␤-activated endothelial cells.…”

Section: Discussionmentioning

confidence: 49%

Short-Term Heat Exposure Inhibits Inflammation by Abrogating Recruitment of and Nuclear Factor-κB Activation in Neutrophils Exposed to Chemotactic Cytokines

Choi

Salanova

Rolle

et al. 2008

The American Journal of Pathology

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Cytokines, such as granulocyte macrophage-colony stimulating factor (GM-CSF) and interleukin (IL)-8 attract neutrophils into inflammatory sites. During emigration from the blood neutrophils interact with extracellular matrix proteins such as fibronectin. Fibronectin provides ␤2-integrin co-stimulation, allowing GM-CSF and IL-8 to activate nuclear factor (NF)-B, an effect that does not occur in suspension. We tested the hypothesis that exposure of mice to fever-like temperatures abrogates neutrophil recruitment and NF-B activation in a mouse model of skin inflammation. Mice that were exposed to 40°C for 1 hour showed strongly reduced GM-CSF-and IL-8-induced neutrophilic skin inflammation. In vitro heat exposure did not interfere with neutrophil adhesion or spreading on fibronectin but strongly inhibited migration toward both cytokines. Using specific inhibitors, we found that PI3-K/Akt was pivotal for neutrophil migration and that heat down-regulated this pathway. Furthermore, neutrophils on fibronectin showed abrogated NF-B activation in response to GM-CSF and IL-8 after heat. In vivo heat exposure of mice followed by ex vivo stimulation of isolated bone marrow neutrophils confirmed these results. Finally, less NF-B activation was seen in the inflammatory lesions of mice exposed to fever-like temperatures as demonstrated by in situ hybridization for IB␣ mRNA. These new findings suggest that heat may have anti-inflammatory effects in neutrophil-dependent inflammation. (Am J Pathol

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Section: Discussionmentioning

confidence: 49%

Short-Term Heat Exposure Inhibits Inflammation by Abrogating Recruitment of and Nuclear Factor-κB Activation in Neutrophils Exposed to Chemotactic Cytokines

Choi

Salanova

Rolle

et al. 2008

The American Journal of Pathology

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“…Recombinant human IL-8 that was used in the crystallographic studies was purified from Escherichia coli (18). All of the x-ray data used for this structure determination were collected on crystals that were grown using protein recycled from the NMR experiments of Clore and Gronenborn and coworkers (6,7).…”

Section: Methodsmentioning

confidence: 99%

Crystal structure of interleukin 8: symbiosis of NMR and crystallography.

Baldwin

Weber

Charles

et al. 1991

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

342

223

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The crystal structure of a host defense system chemotactic factor, interleukin 8, has been solved by molecular replacement using as a model the solution structure derived from nuclear magnetic resonance experiments. The structure was refined with 2 A x-ray data to an R factor of 0.187 (0.217 at 1.6 A). A comparison indicates some potential differences between the structure in solution and in the crystalline state. Our analysis also predicts that residues 4 through 9 on the amino terminus and the (-bend, which includes His-33, may be important for receptor binding.Nuclear magnetic resonance (NMR) has been established in the last 5 years as the only viable alternative to x-ray crystallography for determining detailed three-dimensional structures of macromolecules up to 20 kDa molecular mass (1, 2). Although test calculations using crambin (3), tendamistat (4), and ubiquitin (5) have established the feasibility of utilizing NMR-derived models as starting points for solving x-ray structures by molecular replacement, no unknown crystal structures have been previously solved by that route. We present here the results of such an investigation. The highresolution x-ray structure ofinterleukin 8 (IL-8) was solved by a combination of NMR (6, 7) and crystallography, and the models obtained by these two techniques were compared.tt IL-8 is a small, chemotactic factor released by monocytes and a number of other cell types in response to inflammatory stimuli (8). The factor promotes the directed migration of neutrophils, basophils, and T cells (9) along a concentration gradient to the inflammatory focus. IL-8 has also been reported to initiate the oxidative burst response in neutrophils. The factor has been known as monocyte-derived neutrophil chemotactic factor (10) or neutrophil-activating protein (11,12) among others, but it is now generally designated IL-8 (9). IL-8 is a member of a family of similar immune system proteins including the monocyte chemoattractant factor (13), human melanoma growth-stimulating activity (14), murine macrophage inflammatory protein 2 (15), and bovine platelet factor 4 (BPF4) (16). Each of these protein sequences shows a conserved region containing four cysteines that form two disulfide bridges. Under the experimental conditions of the structural investigations IL-8 is a dimer (6, 7) and BPF4 is a tetramer (17). Materials and MethodsRecombinant human IL-8 that was used in the crystallographic studies was purified from Escherichia coli (18). All of the x-ray data used for this structure determination were collected on crystals that were grown using protein recycled from the NMR experiments of Clore and Gronenborn and coworkers (6, 7). Crystals belonging to the space group P3121, a = b = 40.3 A, c = 90.1 A, were grownfrom40%o ammonium sulfate (pH = 8.5) as described (19). Identical crystals have also been grown using chemically synthesized protein. Recently, workers at Sandoz Pharmaceutical have reported that crystals apparently identical to the ones described here could be grown from poly...

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“…The proinflammatory cytokine interleukin 8/neutrophil-ac-tivating peptide-1 (IL-8/NAP-1)1 (2)(3)(4)(5)(6)(7), has been implicated as one important mediator of neutrophil infiltrates identified in the epidermal scales of psoriasis patients (8,9), the synovial fluid of rheumatoid arthritis patients (10,11), the colonic mucosa of individuals with ulcerative colitis (12,13), and in the lung of patients with idiopathic pulmonary fibrosis or adult respiratory distress syndrome (14, 15 (4). In addition to its proinflammatory role in recruiting neutrophils to local sites of acute and chronic inflammation, IL-8 may have some anti-inflammatory properties.…”

Section: Introductionmentioning

confidence: 99%

Long-term impaired neutrophil migration in mice overexpressing human interleukin-8.

Simonet

Hughes²,

Nguyen³

et al. 1994

J. Clin. Invest.

158

110

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The proinflammatory chemokine interleukin-8 (IL-8/NAP-1) has been implicated in recruiting neutrophils to sites of acute and chronic tissue inflammation. In transgenic mice, elevated serum IL-8 levels ranging from 1 to 118 ng/ml were correlated with proportional increases in circulating neutrophils and proportional decreases in L-selectin expression on the surface of blood neutrophils. No change in the expression of the fi2-integrins Mac-i and LFA-1 was apparent on peripheral blood neutrophils of the IL-8 transgenic mice. Additionally, L-selectin expression on bone marrow neutrophils and neutrophil precursors was normal in all transgenic lines. IL-8 transgenic mice demonstrated an accumulation of neutrophils in the microcirculation of the lung, liver and spleen. Moreover, there was no evidence of neutrophil extravasation, plasma exudation or tissue damage in any IL-8 transgenic mice. Neutrophil migration into the inflamed peritoneal cavity was severely inhibited in IL-8 transgenic mice, but not in nontransgenic littermates. The IL-8 transgenic mice should serve as useful models for studying the putative role of neutrophils in mediating tissue damage in models of inflammation, such as hepatic ischemia and reperfusion injury, cecal puncture and ligation, and glomerulonephritis. (J. Clin. Invest. 1994. 94:1310-1319

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Production and Characterization of Recombinant Human Neutrophil Chemotactic Factor

Cited by 63 publications

References 0 publications

Short-Term Heat Exposure Inhibits Inflammation by Abrogating Recruitment of and Nuclear Factor-κB Activation in Neutrophils Exposed to Chemotactic Cytokines

Short-Term Heat Exposure Inhibits Inflammation by Abrogating Recruitment of and Nuclear Factor-κB Activation in Neutrophils Exposed to Chemotactic Cytokines

Crystal structure of interleukin 8: symbiosis of NMR and crystallography.

Long-term impaired neutrophil migration in mice overexpressing human interleukin-8.

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