A revised model for the secretion of tPA and cytokines from cultured endothelial cells

Laura Knipe, Athinoula Meli, Lindsay Hewlett, Ruben Bierings, John Dempster, Paul Skehel, Matthew J Hannah, Tom Carter

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

Endothelial cells are reported to contain several distinct populations of regulated secretory organelles, including Weibel-Palade bodies (WPBs), the tissue plasminogen activator (tPA) organelle, and the type-2 chemokine-containing organelle. We show that the tPA and type-2 organelles in human endothelial cells represent a single compartment primarily responsible for unstimulated secretion of tPA or, in cells exposed to interleukin-1β (IL-1β), the cytokines IL-8, IL-6, monocyte chemoattractant protein-1 (MCP-1), and growth-regulated oncogene-α (GRO-α). This compartment was distinct from WPBs in that it lacked detectable von Willebrand factor, P-selectin, Rab27a, or CD63 immunoreactivity, displayed no time-dependent decrease in intragranule pH, underwent detectable unstimulated exocytosis, and was very poorly responsive to Ca(2+)-elevating secretagogues. WPBs could also contain tPA, and in IL-1β-treated cells, IL-8, IL-6, MCP-1, and GRO-α, and were the primary source for histamine or ionomycin-stimulated secretion of these molecules. However, analysis of the storage efficiency of cytokines and tPA revealed that all were very poorly stored compared with von Willebrand factor. The nonmammalian, nonsecretory protein EGFP, when expressed in the secretory pathway, also entered WPBs and had a storage efficiency similar to tPA and the cytokines tested. Based on these data, we proposed a revised model for storage and secretion of cytokines and tPA.
LanguageEnglish
Pages2183-2191
Number of pages9
JournalBlood
Volume116
Issue number12
DOIs
Publication statusPublished - 23 Sep 2010

Fingerprint

Endothelial cells
Tissue Plasminogen Activator
Weibel-Palade Bodies
Cultured Cells
Endothelial Cells
Cytokines
Organelles
Chemokine CCL2
von Willebrand Factor
Interleukin-8
Interleukin-1beta
Oncogenes
Interleukin-6
Ionomycin
P-Selectin
Secretory Pathway
Exocytosis
Growth
Chemokines
Histamine

Keywords

  • cell compartmentation
  • cells, cultured
  • cytokines
  • endothelial cells
  • endothelium, vascular
  • humans
  • models, biological
  • tissue plasminogen activator
  • Weibel-Palade bodies

Cite this

Knipe, L., Meli, A., Hewlett, L., Bierings, R., Dempster, J., Skehel, P., ... Carter, T. (2010). A revised model for the secretion of tPA and cytokines from cultured endothelial cells. Blood, 116(12), 2183-2191. https://doi.org/10.1182/blood-2010-03-276170
Knipe, Laura ; Meli, Athinoula ; Hewlett, Lindsay ; Bierings, Ruben ; Dempster, John ; Skehel, Paul ; Hannah, Matthew J ; Carter, Tom. / A revised model for the secretion of tPA and cytokines from cultured endothelial cells. In: Blood. 2010 ; Vol. 116, No. 12. pp. 2183-2191.
@article{0187d918261f4f3e96dfbaa722d3186a,
title = "A revised model for the secretion of tPA and cytokines from cultured endothelial cells",
abstract = "Endothelial cells are reported to contain several distinct populations of regulated secretory organelles, including Weibel-Palade bodies (WPBs), the tissue plasminogen activator (tPA) organelle, and the type-2 chemokine-containing organelle. We show that the tPA and type-2 organelles in human endothelial cells represent a single compartment primarily responsible for unstimulated secretion of tPA or, in cells exposed to interleukin-1β (IL-1β), the cytokines IL-8, IL-6, monocyte chemoattractant protein-1 (MCP-1), and growth-regulated oncogene-α (GRO-α). This compartment was distinct from WPBs in that it lacked detectable von Willebrand factor, P-selectin, Rab27a, or CD63 immunoreactivity, displayed no time-dependent decrease in intragranule pH, underwent detectable unstimulated exocytosis, and was very poorly responsive to Ca(2+)-elevating secretagogues. WPBs could also contain tPA, and in IL-1β-treated cells, IL-8, IL-6, MCP-1, and GRO-α, and were the primary source for histamine or ionomycin-stimulated secretion of these molecules. However, analysis of the storage efficiency of cytokines and tPA revealed that all were very poorly stored compared with von Willebrand factor. The nonmammalian, nonsecretory protein EGFP, when expressed in the secretory pathway, also entered WPBs and had a storage efficiency similar to tPA and the cytokines tested. Based on these data, we proposed a revised model for storage and secretion of cytokines and tPA.",
keywords = "cell compartmentation, cells, cultured, cytokines, endothelial cells, endothelium, vascular, humans, models, biological, tissue plasminogen activator, Weibel-Palade bodies",
author = "Laura Knipe and Athinoula Meli and Lindsay Hewlett and Ruben Bierings and John Dempster and Paul Skehel and Hannah, {Matthew J} and Tom Carter",
year = "2010",
month = "9",
day = "23",
doi = "10.1182/blood-2010-03-276170",
language = "English",
volume = "116",
pages = "2183--2191",
journal = "Blood",
issn = "0006-4971",
number = "12",

}

Knipe, L, Meli, A, Hewlett, L, Bierings, R, Dempster, J, Skehel, P, Hannah, MJ & Carter, T 2010, 'A revised model for the secretion of tPA and cytokines from cultured endothelial cells' Blood, vol. 116, no. 12, pp. 2183-2191. https://doi.org/10.1182/blood-2010-03-276170

A revised model for the secretion of tPA and cytokines from cultured endothelial cells. / Knipe, Laura; Meli, Athinoula; Hewlett, Lindsay; Bierings, Ruben; Dempster, John; Skehel, Paul; Hannah, Matthew J; Carter, Tom.

In: Blood, Vol. 116, No. 12, 23.09.2010, p. 2183-2191.

Research output: Contribution to journalArticle

TY - JOUR

T1 - A revised model for the secretion of tPA and cytokines from cultured endothelial cells

AU - Knipe, Laura

AU - Meli, Athinoula

AU - Hewlett, Lindsay

AU - Bierings, Ruben

AU - Dempster, John

AU - Skehel, Paul

AU - Hannah, Matthew J

AU - Carter, Tom

PY - 2010/9/23

Y1 - 2010/9/23

N2 - Endothelial cells are reported to contain several distinct populations of regulated secretory organelles, including Weibel-Palade bodies (WPBs), the tissue plasminogen activator (tPA) organelle, and the type-2 chemokine-containing organelle. We show that the tPA and type-2 organelles in human endothelial cells represent a single compartment primarily responsible for unstimulated secretion of tPA or, in cells exposed to interleukin-1β (IL-1β), the cytokines IL-8, IL-6, monocyte chemoattractant protein-1 (MCP-1), and growth-regulated oncogene-α (GRO-α). This compartment was distinct from WPBs in that it lacked detectable von Willebrand factor, P-selectin, Rab27a, or CD63 immunoreactivity, displayed no time-dependent decrease in intragranule pH, underwent detectable unstimulated exocytosis, and was very poorly responsive to Ca(2+)-elevating secretagogues. WPBs could also contain tPA, and in IL-1β-treated cells, IL-8, IL-6, MCP-1, and GRO-α, and were the primary source for histamine or ionomycin-stimulated secretion of these molecules. However, analysis of the storage efficiency of cytokines and tPA revealed that all were very poorly stored compared with von Willebrand factor. The nonmammalian, nonsecretory protein EGFP, when expressed in the secretory pathway, also entered WPBs and had a storage efficiency similar to tPA and the cytokines tested. Based on these data, we proposed a revised model for storage and secretion of cytokines and tPA.

AB - Endothelial cells are reported to contain several distinct populations of regulated secretory organelles, including Weibel-Palade bodies (WPBs), the tissue plasminogen activator (tPA) organelle, and the type-2 chemokine-containing organelle. We show that the tPA and type-2 organelles in human endothelial cells represent a single compartment primarily responsible for unstimulated secretion of tPA or, in cells exposed to interleukin-1β (IL-1β), the cytokines IL-8, IL-6, monocyte chemoattractant protein-1 (MCP-1), and growth-regulated oncogene-α (GRO-α). This compartment was distinct from WPBs in that it lacked detectable von Willebrand factor, P-selectin, Rab27a, or CD63 immunoreactivity, displayed no time-dependent decrease in intragranule pH, underwent detectable unstimulated exocytosis, and was very poorly responsive to Ca(2+)-elevating secretagogues. WPBs could also contain tPA, and in IL-1β-treated cells, IL-8, IL-6, MCP-1, and GRO-α, and were the primary source for histamine or ionomycin-stimulated secretion of these molecules. However, analysis of the storage efficiency of cytokines and tPA revealed that all were very poorly stored compared with von Willebrand factor. The nonmammalian, nonsecretory protein EGFP, when expressed in the secretory pathway, also entered WPBs and had a storage efficiency similar to tPA and the cytokines tested. Based on these data, we proposed a revised model for storage and secretion of cytokines and tPA.

KW - cell compartmentation

KW - cells, cultured

KW - cytokines

KW - endothelial cells

KW - endothelium, vascular

KW - humans

KW - models, biological

KW - tissue plasminogen activator

KW - Weibel-Palade bodies

U2 - 10.1182/blood-2010-03-276170

DO - 10.1182/blood-2010-03-276170

M3 - Article

VL - 116

SP - 2183

EP - 2191

JO - Blood

T2 - Blood

JF - Blood

SN - 0006-4971

IS - 12

ER -

Knipe L, Meli A, Hewlett L, Bierings R, Dempster J, Skehel P et al. A revised model for the secretion of tPA and cytokines from cultured endothelial cells. Blood. 2010 Sep 23;116(12):2183-2191. https://doi.org/10.1182/blood-2010-03-276170