Activation of Endothelial Cell Phospholipase D by Migrating Neutrophils

Yi Cui, Denis K. English, Rafat A. Siddiqui, Maria Herenyiova, Joe GN Garcia

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Background: Vascular endothelium plays a critical role in regulating the integrity of intercellular adhesive and junctional contacts in response to migrating neutrophils during the inflammatory process. Biochemical responses induced in endothelial cells by adherent, migrating neutrophils are poorly understood. This study was undertaken to explore the possibility that endothelial cell phospholipase D (PLD) is activated when neutrophils migrate through endothelial cell monolayers in response to chemotactic stimuli. Methods: Bovine pulmonary artery endothelial cells (BPAEC) were cultured on polycarbonate filters at the base of inserts in Transwell chambers (Costar, Cambridge, MA) and subsequently labeled with 3H-myristic acid. Human neutrophil migration through BPAEC monolayers was measured in response to a chemoattractant gradient produced by leukotriene B4 (LTB4). After neutrophils were induced to migrate through endothelial monolayers in the presence of 0.1% ethanol, the filters were excised, and cellular lipids were extracted. Levels of labeled phosphatidylethanol (PEt), an index of activation of endothelial cell PLD, were measured in chromatographs of resolved phospholipids. Results: When neutrophils migrated through endothelial monolayers in response to LTB4, endothelial cells accumulated significant levels of PEt, indicating activation of endothelial cell PLD. Kinetic analysis demonstrated that PEt generation closely paralleled chemotactic migration over a 2-hour time period. Direct contact of neutrophils with the endothelium failed to induce PEt formation in the absence of neutrophil chemotaxis, and LTB4 was an ineffective stimulus of endothelial cell PLD activity in the absence of migrating neutrophils. Neutrophil migration-dependent endothelial PLD activation was observed when chemotaxis was induced by an unrelated chemoattractant, serum activated by exposure to Escherichia coli. However, neutrophil migration alone could not account for activation of endothelial cell PLD, since the peptide chemoattractant f-met-leu-phe (FMLP) induced comparable migration of neutrophils through endothelial monolayers but did not induce PEt generation. Conclusions: Our study indicates that chemotactic migration of neutrophils through endothelial monolayers results in endothelial cell PLD activation. This process may amplify both target and effector cell reactivity during the inflammatory response.

Original languageEnglish (US)
Pages (from-to)388-393
Number of pages6
JournalJournal of Investigative Medicine
Volume45
Issue number6
StatePublished - Aug 1997
Externally publishedYes

Fingerprint

Phospholipase D
Endothelial cells
Neutrophils
Endothelial Cells
Chemical activation
Monolayers
Leukotriene B4
Chemotactic Factors
polycarbonate
Chemotaxis
Pulmonary Artery
methionyl-leucyl-phenylalanine
Myristic Acid
Vascular Endothelium
Escherichia coli
Phospholipids
Adhesives
Ethanol
Endothelium
phosphatidylethanol

Keywords

  • Chemotaxis
  • Neutrophil
  • Phospholipase D
  • Signal transduction
  • Vascular endothelium

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Cui, Y., English, D. K., Siddiqui, R. A., Herenyiova, M., & Garcia, J. GN. (1997). Activation of Endothelial Cell Phospholipase D by Migrating Neutrophils. Journal of Investigative Medicine, 45(6), 388-393.

Activation of Endothelial Cell Phospholipase D by Migrating Neutrophils. / Cui, Yi; English, Denis K.; Siddiqui, Rafat A.; Herenyiova, Maria; Garcia, Joe GN.

In: Journal of Investigative Medicine, Vol. 45, No. 6, 08.1997, p. 388-393.

Research output: Contribution to journalArticle

Cui, Y, English, DK, Siddiqui, RA, Herenyiova, M & Garcia, JGN 1997, 'Activation of Endothelial Cell Phospholipase D by Migrating Neutrophils', Journal of Investigative Medicine, vol. 45, no. 6, pp. 388-393.
Cui, Yi ; English, Denis K. ; Siddiqui, Rafat A. ; Herenyiova, Maria ; Garcia, Joe GN. / Activation of Endothelial Cell Phospholipase D by Migrating Neutrophils. In: Journal of Investigative Medicine. 1997 ; Vol. 45, No. 6. pp. 388-393.
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abstract = "Background: Vascular endothelium plays a critical role in regulating the integrity of intercellular adhesive and junctional contacts in response to migrating neutrophils during the inflammatory process. Biochemical responses induced in endothelial cells by adherent, migrating neutrophils are poorly understood. This study was undertaken to explore the possibility that endothelial cell phospholipase D (PLD) is activated when neutrophils migrate through endothelial cell monolayers in response to chemotactic stimuli. Methods: Bovine pulmonary artery endothelial cells (BPAEC) were cultured on polycarbonate filters at the base of inserts in Transwell chambers (Costar, Cambridge, MA) and subsequently labeled with 3H-myristic acid. Human neutrophil migration through BPAEC monolayers was measured in response to a chemoattractant gradient produced by leukotriene B4 (LTB4). After neutrophils were induced to migrate through endothelial monolayers in the presence of 0.1{\%} ethanol, the filters were excised, and cellular lipids were extracted. Levels of labeled phosphatidylethanol (PEt), an index of activation of endothelial cell PLD, were measured in chromatographs of resolved phospholipids. Results: When neutrophils migrated through endothelial monolayers in response to LTB4, endothelial cells accumulated significant levels of PEt, indicating activation of endothelial cell PLD. Kinetic analysis demonstrated that PEt generation closely paralleled chemotactic migration over a 2-hour time period. Direct contact of neutrophils with the endothelium failed to induce PEt formation in the absence of neutrophil chemotaxis, and LTB4 was an ineffective stimulus of endothelial cell PLD activity in the absence of migrating neutrophils. Neutrophil migration-dependent endothelial PLD activation was observed when chemotaxis was induced by an unrelated chemoattractant, serum activated by exposure to Escherichia coli. However, neutrophil migration alone could not account for activation of endothelial cell PLD, since the peptide chemoattractant f-met-leu-phe (FMLP) induced comparable migration of neutrophils through endothelial monolayers but did not induce PEt generation. Conclusions: Our study indicates that chemotactic migration of neutrophils through endothelial monolayers results in endothelial cell PLD activation. This process may amplify both target and effector cell reactivity during the inflammatory response.",
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