The role of tyrosine phosphorylation in lipopolysaccharide-and zymosan- induced procoagulant activity and tissue factor expression in macrophages

A. P B Dackiw, S. Grinstein, G. F. Brisseau, I. D. McGilvray, A. B. Nathens, J. A. Mcguire, R. Romanek, P. Y C Cheung, O. D. Rotstein

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

The expression of surface procoagulants by exudative macrophages represents an important mechanism underlying local fibrin deposition at sites of extravascular inflammation. The present studies investigated the contribution of tyrosine phosphorylation to the generation of macrophage procoagulant activity (PCA) and tissue factor expression in response to proinflammatory stimuli. Both lipopolysaccharide (LPS) and zymosan rapidly stimulated tyrosine phosphorylation in elicited murine peritoneal macrophages. This effect was prevented by the tyrosine kinase inhibitors genistein and herbimycin and augmented by the addition of the phosphotyrosine phosphatase inhibitor vanadate. The vanadate-mediated rise in phosphotyrosine accumulation was abrogated by the use of diphenylene iodonium, an inhibitor of the respiratory burst oxidase, suggesting a role for peroxides of vanadate as contributors to the tyrosine phosphorylation. This notion was supported by the finding that vanadyl hydroperoxide markedly increased the accumulation of phosphotyrosine residues. To define the role of tyrosine phosphorylation in the induction of macrophage PCA by LPS, the effects of tyrosine kinase inhibition by genistein and herbimycin were investigated. Both agents inhibited the expression of macrophage PCA. Further, Northern blot analysis with the cDNA probe for murine tissue factor indicated that the inhibition occurred at the mRNA level or earlier. Since vanadate augmented phosphotyrosine accumulation, it was hypothesized that it might enhance generation of macrophage products. However, vanadate reduced induction of PCA in response to LPS. By contrast, vanadate augmented basal prostaglandin E2 (PGE2) release and stimulated PGE2 release by macrophages. Indomethacin prevented the increase in PGE2 but only partially restored normal levels of PCA. The effect of vanadate on tissue factor expression appeared to be posttranscriptional. These studies thus demonstrate, by functional Western blotting and Northern blotting techniques, that tyrosine phosphorylation plays a role in the regulation of macrophage PCA and tissue factor expression in response to proinflammatory stimuli.

Original languageEnglish (US)
Pages (from-to)2362-2370
Number of pages9
JournalInfection and Immunity
Volume65
Issue number6
StatePublished - 1997

Fingerprint

Zymosan
Vanadates
Thromboplastin
Tyrosine
Lipopolysaccharides
Macrophages
Phosphorylation
Phosphotyrosine
Dinoprostone
Genistein
Northern Blotting
Protein-Tyrosine Kinases
Protein Tyrosine Phosphatases
Peritoneal Macrophages
Fibrin
Indomethacin
Complementary DNA
Western Blotting
Inflammation
Messenger RNA

ASJC Scopus subject areas

  • Immunology

Cite this

Dackiw, A. P. B., Grinstein, S., Brisseau, G. F., McGilvray, I. D., Nathens, A. B., Mcguire, J. A., ... Rotstein, O. D. (1997). The role of tyrosine phosphorylation in lipopolysaccharide-and zymosan- induced procoagulant activity and tissue factor expression in macrophages. Infection and Immunity, 65(6), 2362-2370.

The role of tyrosine phosphorylation in lipopolysaccharide-and zymosan- induced procoagulant activity and tissue factor expression in macrophages. / Dackiw, A. P B; Grinstein, S.; Brisseau, G. F.; McGilvray, I. D.; Nathens, A. B.; Mcguire, J. A.; Romanek, R.; Cheung, P. Y C; Rotstein, O. D.

In: Infection and Immunity, Vol. 65, No. 6, 1997, p. 2362-2370.

Research output: Contribution to journalArticle

Dackiw, APB, Grinstein, S, Brisseau, GF, McGilvray, ID, Nathens, AB, Mcguire, JA, Romanek, R, Cheung, PYC & Rotstein, OD 1997, 'The role of tyrosine phosphorylation in lipopolysaccharide-and zymosan- induced procoagulant activity and tissue factor expression in macrophages', Infection and Immunity, vol. 65, no. 6, pp. 2362-2370.
Dackiw, A. P B ; Grinstein, S. ; Brisseau, G. F. ; McGilvray, I. D. ; Nathens, A. B. ; Mcguire, J. A. ; Romanek, R. ; Cheung, P. Y C ; Rotstein, O. D. / The role of tyrosine phosphorylation in lipopolysaccharide-and zymosan- induced procoagulant activity and tissue factor expression in macrophages. In: Infection and Immunity. 1997 ; Vol. 65, No. 6. pp. 2362-2370.
@article{55c1f2f456724d348182da4904638859,
title = "The role of tyrosine phosphorylation in lipopolysaccharide-and zymosan- induced procoagulant activity and tissue factor expression in macrophages",
abstract = "The expression of surface procoagulants by exudative macrophages represents an important mechanism underlying local fibrin deposition at sites of extravascular inflammation. The present studies investigated the contribution of tyrosine phosphorylation to the generation of macrophage procoagulant activity (PCA) and tissue factor expression in response to proinflammatory stimuli. Both lipopolysaccharide (LPS) and zymosan rapidly stimulated tyrosine phosphorylation in elicited murine peritoneal macrophages. This effect was prevented by the tyrosine kinase inhibitors genistein and herbimycin and augmented by the addition of the phosphotyrosine phosphatase inhibitor vanadate. The vanadate-mediated rise in phosphotyrosine accumulation was abrogated by the use of diphenylene iodonium, an inhibitor of the respiratory burst oxidase, suggesting a role for peroxides of vanadate as contributors to the tyrosine phosphorylation. This notion was supported by the finding that vanadyl hydroperoxide markedly increased the accumulation of phosphotyrosine residues. To define the role of tyrosine phosphorylation in the induction of macrophage PCA by LPS, the effects of tyrosine kinase inhibition by genistein and herbimycin were investigated. Both agents inhibited the expression of macrophage PCA. Further, Northern blot analysis with the cDNA probe for murine tissue factor indicated that the inhibition occurred at the mRNA level or earlier. Since vanadate augmented phosphotyrosine accumulation, it was hypothesized that it might enhance generation of macrophage products. However, vanadate reduced induction of PCA in response to LPS. By contrast, vanadate augmented basal prostaglandin E2 (PGE2) release and stimulated PGE2 release by macrophages. Indomethacin prevented the increase in PGE2 but only partially restored normal levels of PCA. The effect of vanadate on tissue factor expression appeared to be posttranscriptional. These studies thus demonstrate, by functional Western blotting and Northern blotting techniques, that tyrosine phosphorylation plays a role in the regulation of macrophage PCA and tissue factor expression in response to proinflammatory stimuli.",
author = "Dackiw, {A. P B} and S. Grinstein and Brisseau, {G. F.} and McGilvray, {I. D.} and Nathens, {A. B.} and Mcguire, {J. A.} and R. Romanek and Cheung, {P. Y C} and Rotstein, {O. D.}",
year = "1997",
language = "English (US)",
volume = "65",
pages = "2362--2370",
journal = "Infection and Immunity",
issn = "0019-9567",
publisher = "American Society for Microbiology",
number = "6",

}

TY - JOUR

T1 - The role of tyrosine phosphorylation in lipopolysaccharide-and zymosan- induced procoagulant activity and tissue factor expression in macrophages

AU - Dackiw, A. P B

AU - Grinstein, S.

AU - Brisseau, G. F.

AU - McGilvray, I. D.

AU - Nathens, A. B.

AU - Mcguire, J. A.

AU - Romanek, R.

AU - Cheung, P. Y C

AU - Rotstein, O. D.

PY - 1997

Y1 - 1997

N2 - The expression of surface procoagulants by exudative macrophages represents an important mechanism underlying local fibrin deposition at sites of extravascular inflammation. The present studies investigated the contribution of tyrosine phosphorylation to the generation of macrophage procoagulant activity (PCA) and tissue factor expression in response to proinflammatory stimuli. Both lipopolysaccharide (LPS) and zymosan rapidly stimulated tyrosine phosphorylation in elicited murine peritoneal macrophages. This effect was prevented by the tyrosine kinase inhibitors genistein and herbimycin and augmented by the addition of the phosphotyrosine phosphatase inhibitor vanadate. The vanadate-mediated rise in phosphotyrosine accumulation was abrogated by the use of diphenylene iodonium, an inhibitor of the respiratory burst oxidase, suggesting a role for peroxides of vanadate as contributors to the tyrosine phosphorylation. This notion was supported by the finding that vanadyl hydroperoxide markedly increased the accumulation of phosphotyrosine residues. To define the role of tyrosine phosphorylation in the induction of macrophage PCA by LPS, the effects of tyrosine kinase inhibition by genistein and herbimycin were investigated. Both agents inhibited the expression of macrophage PCA. Further, Northern blot analysis with the cDNA probe for murine tissue factor indicated that the inhibition occurred at the mRNA level or earlier. Since vanadate augmented phosphotyrosine accumulation, it was hypothesized that it might enhance generation of macrophage products. However, vanadate reduced induction of PCA in response to LPS. By contrast, vanadate augmented basal prostaglandin E2 (PGE2) release and stimulated PGE2 release by macrophages. Indomethacin prevented the increase in PGE2 but only partially restored normal levels of PCA. The effect of vanadate on tissue factor expression appeared to be posttranscriptional. These studies thus demonstrate, by functional Western blotting and Northern blotting techniques, that tyrosine phosphorylation plays a role in the regulation of macrophage PCA and tissue factor expression in response to proinflammatory stimuli.

AB - The expression of surface procoagulants by exudative macrophages represents an important mechanism underlying local fibrin deposition at sites of extravascular inflammation. The present studies investigated the contribution of tyrosine phosphorylation to the generation of macrophage procoagulant activity (PCA) and tissue factor expression in response to proinflammatory stimuli. Both lipopolysaccharide (LPS) and zymosan rapidly stimulated tyrosine phosphorylation in elicited murine peritoneal macrophages. This effect was prevented by the tyrosine kinase inhibitors genistein and herbimycin and augmented by the addition of the phosphotyrosine phosphatase inhibitor vanadate. The vanadate-mediated rise in phosphotyrosine accumulation was abrogated by the use of diphenylene iodonium, an inhibitor of the respiratory burst oxidase, suggesting a role for peroxides of vanadate as contributors to the tyrosine phosphorylation. This notion was supported by the finding that vanadyl hydroperoxide markedly increased the accumulation of phosphotyrosine residues. To define the role of tyrosine phosphorylation in the induction of macrophage PCA by LPS, the effects of tyrosine kinase inhibition by genistein and herbimycin were investigated. Both agents inhibited the expression of macrophage PCA. Further, Northern blot analysis with the cDNA probe for murine tissue factor indicated that the inhibition occurred at the mRNA level or earlier. Since vanadate augmented phosphotyrosine accumulation, it was hypothesized that it might enhance generation of macrophage products. However, vanadate reduced induction of PCA in response to LPS. By contrast, vanadate augmented basal prostaglandin E2 (PGE2) release and stimulated PGE2 release by macrophages. Indomethacin prevented the increase in PGE2 but only partially restored normal levels of PCA. The effect of vanadate on tissue factor expression appeared to be posttranscriptional. These studies thus demonstrate, by functional Western blotting and Northern blotting techniques, that tyrosine phosphorylation plays a role in the regulation of macrophage PCA and tissue factor expression in response to proinflammatory stimuli.

UR - http://www.scopus.com/inward/record.url?scp=0030999487&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030999487&partnerID=8YFLogxK

M3 - Article

VL - 65

SP - 2362

EP - 2370

JO - Infection and Immunity

JF - Infection and Immunity

SN - 0019-9567

IS - 6

ER -