Human polymorphonuclear cell death after exposure to resuscitation fluids in vitro: Apoptosis versus necrosis

Kathleen Stanton, Hasan B. Alam, Peter M Rhee, Orlando Llorente, John Kirkpatrick, Elena Koustova, David B. Hoyt, Charles E. Lucas

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Background: Resuscitation fluids can have variable effects on key functions of circulating polymorphonuclear neutrophils (PMNs) such as oxidative burst, chemotaxis, and bacterial killing. We hypothesized that choice of resuscitation fluids will also affect the rate of PMN apoptosis. To test this, we studied cellular death (apoptosis and necrosis) in human PMNs after brief exposure to different hypertonic and isotonic fluids. Methods: Blood from 12 volunteers was incubated for 1 hour at 37°C in normal saline, 6.0% dextran-70, 7.5% hypertonic saline, and 7.5% hypertonic saline with 6% dextran-70. Isolated PMNs were double labeled with fluorescein-Annexin V and propidium iodide, and apoptosis and necrosis were measured using flow cytometry. Caspase activation was also detected with flow cytometry using pancaspase inhibitor (carbobenzoxy-valyl-alanyl-aspartyl-fluoromethylketone) in non-isolated whole blood samples to determine apoptosis. Finally, cDNA macroarrays were used to evaluate the expression of 23 genes involved in the regulation of cell cycling and apoptosis. Results: Exposure to hypertonic fluids (hypertonic saline and 7.5% hypertonic saline with 6% dextran-70) significantly (p < 0.05) increased necrosis in isolated PMNs. In whole blood samples, PMNs exposed to dextran demonstrated significant apoptosis as evidenced by increased caspase activation. Dextran was the only fluid that affected leukocyte gene expression, inducing significant up-regulation of Rb gene transcription. Conclusion: Hypertonic fluids and dextran decrease human polymorphonuclear cell survival through necrotic and apoptotic pathways, respectively.

Original languageEnglish (US)
Pages (from-to)1065-1076
Number of pages12
JournalJournal of Trauma
Volume54
Issue number6
StatePublished - Jun 1 2003
Externally publishedYes

Fingerprint

Dextrans
Resuscitation
Neutrophils
Cell Death
Necrosis
Apoptosis
Caspases
Flow Cytometry
Retinoblastoma Genes
Gene Expression
Respiratory Burst
Propidium
Annexin A5
Chemotaxis
Fluorescein
In Vitro Techniques
Volunteers
Cell Survival
Leukocytes
Up-Regulation

Keywords

  • Apoptosis
  • Cell death
  • Dextran
  • Hypertonic fluids
  • Leukocytes
  • Necrosis
  • Polymorphonuclear neutrophils
  • Resuscitation fluids

ASJC Scopus subject areas

  • Surgery

Cite this

Stanton, K., Alam, H. B., Rhee, P. M., Llorente, O., Kirkpatrick, J., Koustova, E., ... Lucas, C. E. (2003). Human polymorphonuclear cell death after exposure to resuscitation fluids in vitro: Apoptosis versus necrosis. Journal of Trauma, 54(6), 1065-1076.

Human polymorphonuclear cell death after exposure to resuscitation fluids in vitro : Apoptosis versus necrosis. / Stanton, Kathleen; Alam, Hasan B.; Rhee, Peter M; Llorente, Orlando; Kirkpatrick, John; Koustova, Elena; Hoyt, David B.; Lucas, Charles E.

In: Journal of Trauma, Vol. 54, No. 6, 01.06.2003, p. 1065-1076.

Research output: Contribution to journalArticle

Stanton, K, Alam, HB, Rhee, PM, Llorente, O, Kirkpatrick, J, Koustova, E, Hoyt, DB & Lucas, CE 2003, 'Human polymorphonuclear cell death after exposure to resuscitation fluids in vitro: Apoptosis versus necrosis', Journal of Trauma, vol. 54, no. 6, pp. 1065-1076.
Stanton K, Alam HB, Rhee PM, Llorente O, Kirkpatrick J, Koustova E et al. Human polymorphonuclear cell death after exposure to resuscitation fluids in vitro: Apoptosis versus necrosis. Journal of Trauma. 2003 Jun 1;54(6):1065-1076.
Stanton, Kathleen ; Alam, Hasan B. ; Rhee, Peter M ; Llorente, Orlando ; Kirkpatrick, John ; Koustova, Elena ; Hoyt, David B. ; Lucas, Charles E. / Human polymorphonuclear cell death after exposure to resuscitation fluids in vitro : Apoptosis versus necrosis. In: Journal of Trauma. 2003 ; Vol. 54, No. 6. pp. 1065-1076.
@article{5731eb9132fe410ba23ccc65600e215d,
title = "Human polymorphonuclear cell death after exposure to resuscitation fluids in vitro: Apoptosis versus necrosis",
abstract = "Background: Resuscitation fluids can have variable effects on key functions of circulating polymorphonuclear neutrophils (PMNs) such as oxidative burst, chemotaxis, and bacterial killing. We hypothesized that choice of resuscitation fluids will also affect the rate of PMN apoptosis. To test this, we studied cellular death (apoptosis and necrosis) in human PMNs after brief exposure to different hypertonic and isotonic fluids. Methods: Blood from 12 volunteers was incubated for 1 hour at 37°C in normal saline, 6.0{\%} dextran-70, 7.5{\%} hypertonic saline, and 7.5{\%} hypertonic saline with 6{\%} dextran-70. Isolated PMNs were double labeled with fluorescein-Annexin V and propidium iodide, and apoptosis and necrosis were measured using flow cytometry. Caspase activation was also detected with flow cytometry using pancaspase inhibitor (carbobenzoxy-valyl-alanyl-aspartyl-fluoromethylketone) in non-isolated whole blood samples to determine apoptosis. Finally, cDNA macroarrays were used to evaluate the expression of 23 genes involved in the regulation of cell cycling and apoptosis. Results: Exposure to hypertonic fluids (hypertonic saline and 7.5{\%} hypertonic saline with 6{\%} dextran-70) significantly (p < 0.05) increased necrosis in isolated PMNs. In whole blood samples, PMNs exposed to dextran demonstrated significant apoptosis as evidenced by increased caspase activation. Dextran was the only fluid that affected leukocyte gene expression, inducing significant up-regulation of Rb gene transcription. Conclusion: Hypertonic fluids and dextran decrease human polymorphonuclear cell survival through necrotic and apoptotic pathways, respectively.",
keywords = "Apoptosis, Cell death, Dextran, Hypertonic fluids, Leukocytes, Necrosis, Polymorphonuclear neutrophils, Resuscitation fluids",
author = "Kathleen Stanton and Alam, {Hasan B.} and Rhee, {Peter M} and Orlando Llorente and John Kirkpatrick and Elena Koustova and Hoyt, {David B.} and Lucas, {Charles E.}",
year = "2003",
month = "6",
day = "1",
language = "English (US)",
volume = "54",
pages = "1065--1076",
journal = "Journal of Trauma and Acute Care Surgery",
issn = "2163-0755",
publisher = "Lippincott Williams and Wilkins",
number = "6",

}

TY - JOUR

T1 - Human polymorphonuclear cell death after exposure to resuscitation fluids in vitro

T2 - Apoptosis versus necrosis

AU - Stanton, Kathleen

AU - Alam, Hasan B.

AU - Rhee, Peter M

AU - Llorente, Orlando

AU - Kirkpatrick, John

AU - Koustova, Elena

AU - Hoyt, David B.

AU - Lucas, Charles E.

PY - 2003/6/1

Y1 - 2003/6/1

N2 - Background: Resuscitation fluids can have variable effects on key functions of circulating polymorphonuclear neutrophils (PMNs) such as oxidative burst, chemotaxis, and bacterial killing. We hypothesized that choice of resuscitation fluids will also affect the rate of PMN apoptosis. To test this, we studied cellular death (apoptosis and necrosis) in human PMNs after brief exposure to different hypertonic and isotonic fluids. Methods: Blood from 12 volunteers was incubated for 1 hour at 37°C in normal saline, 6.0% dextran-70, 7.5% hypertonic saline, and 7.5% hypertonic saline with 6% dextran-70. Isolated PMNs were double labeled with fluorescein-Annexin V and propidium iodide, and apoptosis and necrosis were measured using flow cytometry. Caspase activation was also detected with flow cytometry using pancaspase inhibitor (carbobenzoxy-valyl-alanyl-aspartyl-fluoromethylketone) in non-isolated whole blood samples to determine apoptosis. Finally, cDNA macroarrays were used to evaluate the expression of 23 genes involved in the regulation of cell cycling and apoptosis. Results: Exposure to hypertonic fluids (hypertonic saline and 7.5% hypertonic saline with 6% dextran-70) significantly (p < 0.05) increased necrosis in isolated PMNs. In whole blood samples, PMNs exposed to dextran demonstrated significant apoptosis as evidenced by increased caspase activation. Dextran was the only fluid that affected leukocyte gene expression, inducing significant up-regulation of Rb gene transcription. Conclusion: Hypertonic fluids and dextran decrease human polymorphonuclear cell survival through necrotic and apoptotic pathways, respectively.

AB - Background: Resuscitation fluids can have variable effects on key functions of circulating polymorphonuclear neutrophils (PMNs) such as oxidative burst, chemotaxis, and bacterial killing. We hypothesized that choice of resuscitation fluids will also affect the rate of PMN apoptosis. To test this, we studied cellular death (apoptosis and necrosis) in human PMNs after brief exposure to different hypertonic and isotonic fluids. Methods: Blood from 12 volunteers was incubated for 1 hour at 37°C in normal saline, 6.0% dextran-70, 7.5% hypertonic saline, and 7.5% hypertonic saline with 6% dextran-70. Isolated PMNs were double labeled with fluorescein-Annexin V and propidium iodide, and apoptosis and necrosis were measured using flow cytometry. Caspase activation was also detected with flow cytometry using pancaspase inhibitor (carbobenzoxy-valyl-alanyl-aspartyl-fluoromethylketone) in non-isolated whole blood samples to determine apoptosis. Finally, cDNA macroarrays were used to evaluate the expression of 23 genes involved in the regulation of cell cycling and apoptosis. Results: Exposure to hypertonic fluids (hypertonic saline and 7.5% hypertonic saline with 6% dextran-70) significantly (p < 0.05) increased necrosis in isolated PMNs. In whole blood samples, PMNs exposed to dextran demonstrated significant apoptosis as evidenced by increased caspase activation. Dextran was the only fluid that affected leukocyte gene expression, inducing significant up-regulation of Rb gene transcription. Conclusion: Hypertonic fluids and dextran decrease human polymorphonuclear cell survival through necrotic and apoptotic pathways, respectively.

KW - Apoptosis

KW - Cell death

KW - Dextran

KW - Hypertonic fluids

KW - Leukocytes

KW - Necrosis

KW - Polymorphonuclear neutrophils

KW - Resuscitation fluids

UR - http://www.scopus.com/inward/record.url?scp=0037786701&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037786701&partnerID=8YFLogxK

M3 - Article

C2 - 12813324

AN - SCOPUS:0037786701

VL - 54

SP - 1065

EP - 1076

JO - Journal of Trauma and Acute Care Surgery

JF - Journal of Trauma and Acute Care Surgery

SN - 2163-0755

IS - 6

ER -