Effects of E-selectin and P-selectin blockade on neutrophil sequestration in tissues and neutrophil oxidative burst in burned rats

John F. Hansbrough, Thore Wikström, Magnus Braide, Mayer Tenenhaus, Oliver H. Rennekampff, Verena Kiessig, Ramon Zapata Sirvent, Lars M. Bjursten

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Objective: Neutrophil deposition in tissues (leukosequestration) after shock may produce local tissue injury from proteases and high-energy oxygen species released from sequestered neutrophils. The initial step in the binding of neutrophils to capillary endothelium is the interaction of adhesion molecule (selectin) receptors between neutrophils and endothelial cells. We quantified leukosequestration in the tissues of burned rats using two methods of analysis: a) measurement of lung myeloperoxidase; and b) measurement of radiolabeled neutrophils and erythrocytes deposited in multiple tissues. We then determined the ability of a selectin receptor blocking agent to affect neutrophil deposition in tissues after burn injury. Design: Prospective, controlled, laboratory study. Setting: University research laboratory. Subjects: Male Wistar rats (200 to 300 g). Interventions: After tracheostomy and venous cannulation, rats received 17% total body surface area full-thickness contact burns and were resuscitated with saline (20 mL ip). Experimental animals received 2 mg/kg body weight iv administration of a P-and E-selectin blocking monoclonal antibody, CY-1747, immediately after burn. Lung tissue neutrophils were estimated by measuring myeloperoxidase in lung tissue. Neutrophil retention in lung, liver, spleen, gut, skin, muscle, kidney, and brain tissues was determined by removing (preburn) and differentially radiolabeling neutrophils (111In) and erythrocytes (51Cr), reinfusing cells 4.5 hrs after burn, and measuring tissue radioactivity 30 mins later. Edema was estimated by measuring extravasated 125I-labeled albumin in the various tissues. Peripheral blood neutrophils were analyzed for intracellular hydrogen peroxide content, utilizing a fluorescent dye that reacts with hydrogen peroxide, coupled with analysis of cell fluorescence by flow cytometry. Measurements and Main Results: Myeloperoxidase concentration was increased in lungs 5 hrs after burn (p < .05), indicating neutrophil deposition. Radioisotope studies demonstrated significant (p < .05) leukosequestration into the lung, gut, kidney, skin, and brain tissues at 5 hrs after burn. Flow cytometry showed increased intracellular hydrogen peroxide content in peripheral blood neutrophils 5 hrs after burn. Tissue edema, manifested by radiolabeled albumin retention, was not seen in any tissues. Postburn neutrophil deposition in lungs and liver was blocked (p < .05) by administration of CY- 1747 after burn, but maximal neutrophil hydrogen peroxide content was unaffected. Conclusion: Burn injury in rats results in accumulation of neutrophils in multiple tissues. Neutrophil deposition in the lungs and liver is blocked by administration of the E/P-selectin blocking antibody, CY-1747. Since sequestration of metabolically active neutrophils may induce tissue injury, therapies that block postburn leukosequestration may improve clinical outcomes by limiting remote tissue injury.

Original languageEnglish (US)
Pages (from-to)1366-1372
Number of pages7
JournalCritical Care Medicine
Volume24
Issue number8
DOIs
StatePublished - Jan 1 1996
Externally publishedYes

Fingerprint

P-Selectin
E-Selectin
Respiratory Burst
Neutrophils
Lung
Hydrogen Peroxide
Peroxidase
Wounds and Injuries
Selectins
Blocking Antibodies
Albumins
Liver
Edema
Flow Cytometry
Erythrocytes
Kidney
Skin
Tracheostomy
Body Surface Area
Vascular Endothelium

Keywords

  • burn
  • critical injury
  • inflammation
  • lung injury
  • monoclonal antibody
  • myeloperoxidase
  • neutrophil
  • rats
  • selectin
  • tissue injury

ASJC Scopus subject areas

  • Critical Care and Intensive Care Medicine

Cite this

Hansbrough, J. F., Wikström, T., Braide, M., Tenenhaus, M., Rennekampff, O. H., Kiessig, V., ... Bjursten, L. M. (1996). Effects of E-selectin and P-selectin blockade on neutrophil sequestration in tissues and neutrophil oxidative burst in burned rats. Critical Care Medicine, 24(8), 1366-1372. https://doi.org/10.1097/00003246-199608000-00016

Effects of E-selectin and P-selectin blockade on neutrophil sequestration in tissues and neutrophil oxidative burst in burned rats. / Hansbrough, John F.; Wikström, Thore; Braide, Magnus; Tenenhaus, Mayer; Rennekampff, Oliver H.; Kiessig, Verena; Zapata Sirvent, Ramon; Bjursten, Lars M.

In: Critical Care Medicine, Vol. 24, No. 8, 01.01.1996, p. 1366-1372.

Research output: Contribution to journalArticle

Hansbrough, John F. ; Wikström, Thore ; Braide, Magnus ; Tenenhaus, Mayer ; Rennekampff, Oliver H. ; Kiessig, Verena ; Zapata Sirvent, Ramon ; Bjursten, Lars M. / Effects of E-selectin and P-selectin blockade on neutrophil sequestration in tissues and neutrophil oxidative burst in burned rats. In: Critical Care Medicine. 1996 ; Vol. 24, No. 8. pp. 1366-1372.
@article{60c8cd99d9fa4725a11bf18b340862bb,
title = "Effects of E-selectin and P-selectin blockade on neutrophil sequestration in tissues and neutrophil oxidative burst in burned rats",
abstract = "Objective: Neutrophil deposition in tissues (leukosequestration) after shock may produce local tissue injury from proteases and high-energy oxygen species released from sequestered neutrophils. The initial step in the binding of neutrophils to capillary endothelium is the interaction of adhesion molecule (selectin) receptors between neutrophils and endothelial cells. We quantified leukosequestration in the tissues of burned rats using two methods of analysis: a) measurement of lung myeloperoxidase; and b) measurement of radiolabeled neutrophils and erythrocytes deposited in multiple tissues. We then determined the ability of a selectin receptor blocking agent to affect neutrophil deposition in tissues after burn injury. Design: Prospective, controlled, laboratory study. Setting: University research laboratory. Subjects: Male Wistar rats (200 to 300 g). Interventions: After tracheostomy and venous cannulation, rats received 17{\%} total body surface area full-thickness contact burns and were resuscitated with saline (20 mL ip). Experimental animals received 2 mg/kg body weight iv administration of a P-and E-selectin blocking monoclonal antibody, CY-1747, immediately after burn. Lung tissue neutrophils were estimated by measuring myeloperoxidase in lung tissue. Neutrophil retention in lung, liver, spleen, gut, skin, muscle, kidney, and brain tissues was determined by removing (preburn) and differentially radiolabeling neutrophils (111In) and erythrocytes (51Cr), reinfusing cells 4.5 hrs after burn, and measuring tissue radioactivity 30 mins later. Edema was estimated by measuring extravasated 125I-labeled albumin in the various tissues. Peripheral blood neutrophils were analyzed for intracellular hydrogen peroxide content, utilizing a fluorescent dye that reacts with hydrogen peroxide, coupled with analysis of cell fluorescence by flow cytometry. Measurements and Main Results: Myeloperoxidase concentration was increased in lungs 5 hrs after burn (p < .05), indicating neutrophil deposition. Radioisotope studies demonstrated significant (p < .05) leukosequestration into the lung, gut, kidney, skin, and brain tissues at 5 hrs after burn. Flow cytometry showed increased intracellular hydrogen peroxide content in peripheral blood neutrophils 5 hrs after burn. Tissue edema, manifested by radiolabeled albumin retention, was not seen in any tissues. Postburn neutrophil deposition in lungs and liver was blocked (p < .05) by administration of CY- 1747 after burn, but maximal neutrophil hydrogen peroxide content was unaffected. Conclusion: Burn injury in rats results in accumulation of neutrophils in multiple tissues. Neutrophil deposition in the lungs and liver is blocked by administration of the E/P-selectin blocking antibody, CY-1747. Since sequestration of metabolically active neutrophils may induce tissue injury, therapies that block postburn leukosequestration may improve clinical outcomes by limiting remote tissue injury.",
keywords = "burn, critical injury, inflammation, lung injury, monoclonal antibody, myeloperoxidase, neutrophil, rats, selectin, tissue injury",
author = "Hansbrough, {John F.} and Thore Wikstr{\"o}m and Magnus Braide and Mayer Tenenhaus and Rennekampff, {Oliver H.} and Verena Kiessig and {Zapata Sirvent}, Ramon and Bjursten, {Lars M.}",
year = "1996",
month = "1",
day = "1",
doi = "10.1097/00003246-199608000-00016",
language = "English (US)",
volume = "24",
pages = "1366--1372",
journal = "Critical Care Medicine",
issn = "0090-3493",
publisher = "Lippincott Williams and Wilkins",
number = "8",

}

TY - JOUR

T1 - Effects of E-selectin and P-selectin blockade on neutrophil sequestration in tissues and neutrophil oxidative burst in burned rats

AU - Hansbrough, John F.

AU - Wikström, Thore

AU - Braide, Magnus

AU - Tenenhaus, Mayer

AU - Rennekampff, Oliver H.

AU - Kiessig, Verena

AU - Zapata Sirvent, Ramon

AU - Bjursten, Lars M.

PY - 1996/1/1

Y1 - 1996/1/1

N2 - Objective: Neutrophil deposition in tissues (leukosequestration) after shock may produce local tissue injury from proteases and high-energy oxygen species released from sequestered neutrophils. The initial step in the binding of neutrophils to capillary endothelium is the interaction of adhesion molecule (selectin) receptors between neutrophils and endothelial cells. We quantified leukosequestration in the tissues of burned rats using two methods of analysis: a) measurement of lung myeloperoxidase; and b) measurement of radiolabeled neutrophils and erythrocytes deposited in multiple tissues. We then determined the ability of a selectin receptor blocking agent to affect neutrophil deposition in tissues after burn injury. Design: Prospective, controlled, laboratory study. Setting: University research laboratory. Subjects: Male Wistar rats (200 to 300 g). Interventions: After tracheostomy and venous cannulation, rats received 17% total body surface area full-thickness contact burns and were resuscitated with saline (20 mL ip). Experimental animals received 2 mg/kg body weight iv administration of a P-and E-selectin blocking monoclonal antibody, CY-1747, immediately after burn. Lung tissue neutrophils were estimated by measuring myeloperoxidase in lung tissue. Neutrophil retention in lung, liver, spleen, gut, skin, muscle, kidney, and brain tissues was determined by removing (preburn) and differentially radiolabeling neutrophils (111In) and erythrocytes (51Cr), reinfusing cells 4.5 hrs after burn, and measuring tissue radioactivity 30 mins later. Edema was estimated by measuring extravasated 125I-labeled albumin in the various tissues. Peripheral blood neutrophils were analyzed for intracellular hydrogen peroxide content, utilizing a fluorescent dye that reacts with hydrogen peroxide, coupled with analysis of cell fluorescence by flow cytometry. Measurements and Main Results: Myeloperoxidase concentration was increased in lungs 5 hrs after burn (p < .05), indicating neutrophil deposition. Radioisotope studies demonstrated significant (p < .05) leukosequestration into the lung, gut, kidney, skin, and brain tissues at 5 hrs after burn. Flow cytometry showed increased intracellular hydrogen peroxide content in peripheral blood neutrophils 5 hrs after burn. Tissue edema, manifested by radiolabeled albumin retention, was not seen in any tissues. Postburn neutrophil deposition in lungs and liver was blocked (p < .05) by administration of CY- 1747 after burn, but maximal neutrophil hydrogen peroxide content was unaffected. Conclusion: Burn injury in rats results in accumulation of neutrophils in multiple tissues. Neutrophil deposition in the lungs and liver is blocked by administration of the E/P-selectin blocking antibody, CY-1747. Since sequestration of metabolically active neutrophils may induce tissue injury, therapies that block postburn leukosequestration may improve clinical outcomes by limiting remote tissue injury.

AB - Objective: Neutrophil deposition in tissues (leukosequestration) after shock may produce local tissue injury from proteases and high-energy oxygen species released from sequestered neutrophils. The initial step in the binding of neutrophils to capillary endothelium is the interaction of adhesion molecule (selectin) receptors between neutrophils and endothelial cells. We quantified leukosequestration in the tissues of burned rats using two methods of analysis: a) measurement of lung myeloperoxidase; and b) measurement of radiolabeled neutrophils and erythrocytes deposited in multiple tissues. We then determined the ability of a selectin receptor blocking agent to affect neutrophil deposition in tissues after burn injury. Design: Prospective, controlled, laboratory study. Setting: University research laboratory. Subjects: Male Wistar rats (200 to 300 g). Interventions: After tracheostomy and venous cannulation, rats received 17% total body surface area full-thickness contact burns and were resuscitated with saline (20 mL ip). Experimental animals received 2 mg/kg body weight iv administration of a P-and E-selectin blocking monoclonal antibody, CY-1747, immediately after burn. Lung tissue neutrophils were estimated by measuring myeloperoxidase in lung tissue. Neutrophil retention in lung, liver, spleen, gut, skin, muscle, kidney, and brain tissues was determined by removing (preburn) and differentially radiolabeling neutrophils (111In) and erythrocytes (51Cr), reinfusing cells 4.5 hrs after burn, and measuring tissue radioactivity 30 mins later. Edema was estimated by measuring extravasated 125I-labeled albumin in the various tissues. Peripheral blood neutrophils were analyzed for intracellular hydrogen peroxide content, utilizing a fluorescent dye that reacts with hydrogen peroxide, coupled with analysis of cell fluorescence by flow cytometry. Measurements and Main Results: Myeloperoxidase concentration was increased in lungs 5 hrs after burn (p < .05), indicating neutrophil deposition. Radioisotope studies demonstrated significant (p < .05) leukosequestration into the lung, gut, kidney, skin, and brain tissues at 5 hrs after burn. Flow cytometry showed increased intracellular hydrogen peroxide content in peripheral blood neutrophils 5 hrs after burn. Tissue edema, manifested by radiolabeled albumin retention, was not seen in any tissues. Postburn neutrophil deposition in lungs and liver was blocked (p < .05) by administration of CY- 1747 after burn, but maximal neutrophil hydrogen peroxide content was unaffected. Conclusion: Burn injury in rats results in accumulation of neutrophils in multiple tissues. Neutrophil deposition in the lungs and liver is blocked by administration of the E/P-selectin blocking antibody, CY-1747. Since sequestration of metabolically active neutrophils may induce tissue injury, therapies that block postburn leukosequestration may improve clinical outcomes by limiting remote tissue injury.

KW - burn

KW - critical injury

KW - inflammation

KW - lung injury

KW - monoclonal antibody

KW - myeloperoxidase

KW - neutrophil

KW - rats

KW - selectin

KW - tissue injury

UR - http://www.scopus.com/inward/record.url?scp=0029839316&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029839316&partnerID=8YFLogxK

U2 - 10.1097/00003246-199608000-00016

DO - 10.1097/00003246-199608000-00016

M3 - Article

VL - 24

SP - 1366

EP - 1372

JO - Critical Care Medicine

JF - Critical Care Medicine

SN - 0090-3493

IS - 8

ER -