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. 2001 Dec 18;98(26):15143-8.
doi: 10.1073/pnas.191498798. Epub 2001 Dec 11.

Circulating thioredoxin suppresses lipopolysaccharide-induced neutrophil chemotaxis (V体育ios版)

H Nakamura  1 , L A Herzenberg, J Bai, S Araya, N Kondo, Y Nishinaka, L A Herzenberg, J Yodoi
Affiliations

Circulating thioredoxin suppresses lipopolysaccharide-induced neutrophil chemotaxis

H Nakamura et al. Proc Natl Acad Sci U S A. .

Abstract

Thioredoxin (Trx), a redox enzyme with a conserved active site (Cys-32-Gly-Pro-Cys-35), is induced and secreted into circulation in response to inflammation. Studies here demonstrate that elevating Trx levels in circulation either by i. v. injection of recombinant Trx or stimulating Trx release in Trx-transgenic mice dramatically blocks lipopolysaccharide (LPS)-stimulated neutrophil migration in the murine air pouch chemotaxis model. Furthermore, we show that leukocyte recruitment induced by the murine chemokines KC/GROalpha, RANTES (regulated upon activation, normal T cell expressed and secreted), and monocyte chemoattractant protein-1 (MCP-1) is suppressed also in Trx-transgenic mice VSports手机版. Addressing the mechanism responsible for this suppression, we show that circulating Trx blocks (i) the LPS-stimulated in vitro activation of neutrophil p38 mitogen-activated protein kinase, (ii) the normal down-regulation of CD62L on neutrophils migrating into the LPS-stimulated air pouch, and (iii) the in vitro adhesion of LPS-activated neutrophils on endothelial cells. However, as we also show, Trx does not alter the expression of endothelial cell adhesion molecules (intercellular adhesion molecule-1, vascular cell adhesion molecule-1, CD62P, and CD62E) within 3 h. Collectively, these findings indicate that elevated levels of circulating Trx interfere with chemotaxis by acting directly on neutrophils. We discuss these findings in the context of recent studies reporting beneficial effects of acutely elevated Trx in ischemic injury and negative effects associated with chronically elevated Trx in HIV disease. .

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Figures

Figure 1
Figure 1
Four hours after LPS injection in the air pouch, neutrophils predominate among cells recovered from C57BL/6 (wild type, Left), whereas very few cells (2 × 104 cells) were collected after saline injection (Middle). Cells (2 × 105) were recovered from the Trx-transgenic air pouch where lymphocytes predominate among viable cells recovered (Right). FACS analyses are shown for 10,000 cells in each case; forward scatter units are shown for the x axis, and side scatter units are shown for the y axis.
Figure 2
Figure 2
When C57BL/6 mouse was i.v. injected with 40 μg of human recombinant Trx, blood levels of human Trx were measured by ELISA at the indicated times. Similar results were obtained in three mice (Upper Left). Trx blood levels measured by ELISA after i.v. injection of 1 μg of LPS in the air pouch in Trx-transgenic mice reveal rapid release of Trx into circulation (Upper Right). Air pouches were injected with LPS immediately after animals were i.v. injected with the indicated amount of human recombinant Trx. The bars show the numbers of leukocytes recovered from the LPS-injected air pouches 4 h later (Lower Left). Numbers of infiltrated cells induced by LPS were decreased by both reduced and oxidized recombinant wild-type Trx but not C32S/C35S mutant (Lower Right). Reduced recombinant Trx was prepared by incubation with DTT at 37°C for 30 min, and excess DTT was removed by a Sephadex G-25 column (NAP-5 column, Amersham Pharmacia Biotech). Oxidized recombinant Trx was prepared by air-bubbling on ice for 30 min. The redox status of Trx was determined by 5,5′-dithiobis(2-nitrobenzoic acid) assay. The sulfhydryl residues per molecule measured by the DTNB assay were 4.9 and 2.9 in reduced and oxidized Trx, respectively. Oxidized or reduced Trx was i.v. injected just before LPS injection in the air pouch. Four hours later infiltrated cells were collected. Infiltrated cell numbers are shown as the mean ± SD (Lower Right).
Figure 3
Figure 3
Human neutrophils and lymphocytes were preincubated with 10 μg/ml recombinant wild type Trx or C32S/C35S mutant at 37°C for 30 min and then treated with 1 μg/ml LPS at 37°C for 1 h. Recombinant Trx but not C32S/C35S mutant suppresses the phosphorylation of p38 MAPK in LPS-stimulated neutrophils.
Figure 4
Figure 4
CD62L expression on neutrophils found in the air pouch when sterile saline was injected (Bottom) or recruited when LPS was injected into the pouch in C57/BL6 mice (Top). The middle panels show CD62L expression on neutrophils recovered from LPS-injected air pouches in Trx-transgenic mice (Lower Middle) or in mice pretreated by i.v. injection of 40 μg of Trx (Upper Middle). Leukocytes (10,000) gated in a neutrophil region similar to that in Fig. 1 (Left) were analyzed.
Figure 5
Figure 5
Recombinant Trx, but not C32S/C35S mutant Trx, suppresses the adhesion of neutrophils on HUVEC by adhesion assay using 2′,7′-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein acetoxymethyl ester (Left). Shown are photographs of neutrophils adhering on endothelial cells. The adhesion of LPS-activated human neutrophils on HUVECs is inhibited by the administration of Trx (Middle) but not by C32S/C35S mutant Trx (Right).
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"VSports手机版" References

    1. Holmgren A. Annu Rev Biochem. 1985;54:237–271. - PubMed
    1. Chae H Z, Robison K, Poole L B, Church G, Storz G, Rhee S G. Proc Natl Acad Sci USA. 1994;91:7017–7021. - PMC (V体育2025版) - PubMed
    1. Hirota K, Matsui M, Iwata S, Nishiyama A, Mori K, Yodoi J. Proc Natl Acad Sci USA. 1997;94:3633–3638. - VSports最新版本 - PMC - PubMed
    1. Hirota K, Murata M, Sachi Y, Nakamura H, Takeuchi J, Mori K, Yodoi J. J Biol Chem. 1999;274:27891–27897. - PubMed
    1. Makino Y, Okamoto K, Yoshikawa N, Aoshima M, Hirota K, Yodoi J, Umesono K, Makino I, Tanaka H. J Clin Invest. 1996;98:2469–2477. - PMC - PubMed

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