Extracellular Ubiquitin Increases in Packed Red Blood Cell Units During Storage

Mayur B. Patel, Kenneth G Proctor, Matthias Majetschak

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Background: Ubiquitin (Ub) is involved in intracellular protein metabolism, but may also have extracellular roles in host defense and immunomodulation. Erythrocytes contain high amounts of Ub and hemolysis is one potential source of extracellular Ub in vivo. Since hemolysis also occurs with storage of packed RBC units (pRBCs) in vitro, we hypothesized that Ub is released during storage and that it correlates with immunological properties of pRBCs. Materials and methods: Daily aliquots were drawn from pRBCs (n = 3) for 42 days and plasma was isolated. Ub was measured by ELISA. Immunomodulatory properties of plasma were assessed by measuring endotoxin-stimulated cytokine (TNF-α, IL-6, IL-8) production of normal whole blood, and cell proliferation in phytohemagglutinin-stimulated peripheral blood mononuclear cells. Results: Plasma Ub linearly increased (49 ± 2 ng/mL/day; r2 = 0.82, P < 0.001) 20-fold to 2170 ± 268 ng/mL on day 42. Plasma inhibited TNF-α production but stimulated IL-8 production of normal whole blood, which correlated with time-dependent Ub release (TNFα: rspearman = -0.626, P < 0.001; IL-8: rspearman = 0.427, P = 0.004). Addition of exogenous Ub (equaling day 42 concentration) to day 0-4 plasma inhibited TNF-α production by one-third of the effect detected for day 42 plasma, but also inhibited IL-8 production by 40%. IL-6 production and cell proliferation was unchanged between day 0-4 plasma with or without Ub supplementation and day 42 plasma. Conclusions: Extracellular Ub release in pRBCs correlates with in vitro immunomodulatory effects and may partially contribute to transfusion-related immune modulation. Additionally, the linear kinetics of the ubiquitin release during pRBC storage suggest Ub is a suitable in vitro quality control parameter.

Original languageEnglish
Pages (from-to)226-232
Number of pages7
JournalJournal of Surgical Research
Volume135
Issue number2
DOIs
StatePublished - Oct 1 2006

Fingerprint

Ubiquitin
Erythrocytes
Interleukin-8
Hemolysis
Interleukin-6
Blood Cells
Cell Proliferation
Immunomodulation
Phytohemagglutinins
Endotoxins
Quality Control
Enzyme-Linked Immunosorbent Assay
Cytokines

Keywords

  • blood storage
  • extracellular ubiquitin
  • quality control
  • RBCs
  • transfusion

ASJC Scopus subject areas

  • Surgery

Cite this

Extracellular Ubiquitin Increases in Packed Red Blood Cell Units During Storage. / Patel, Mayur B.; Proctor, Kenneth G; Majetschak, Matthias.

In: Journal of Surgical Research, Vol. 135, No. 2, 01.10.2006, p. 226-232.

Research output: Contribution to journalArticle

Patel, Mayur B. ; Proctor, Kenneth G ; Majetschak, Matthias. / Extracellular Ubiquitin Increases in Packed Red Blood Cell Units During Storage. In: Journal of Surgical Research. 2006 ; Vol. 135, No. 2. pp. 226-232.
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abstract = "Background: Ubiquitin (Ub) is involved in intracellular protein metabolism, but may also have extracellular roles in host defense and immunomodulation. Erythrocytes contain high amounts of Ub and hemolysis is one potential source of extracellular Ub in vivo. Since hemolysis also occurs with storage of packed RBC units (pRBCs) in vitro, we hypothesized that Ub is released during storage and that it correlates with immunological properties of pRBCs. Materials and methods: Daily aliquots were drawn from pRBCs (n = 3) for 42 days and plasma was isolated. Ub was measured by ELISA. Immunomodulatory properties of plasma were assessed by measuring endotoxin-stimulated cytokine (TNF-α, IL-6, IL-8) production of normal whole blood, and cell proliferation in phytohemagglutinin-stimulated peripheral blood mononuclear cells. Results: Plasma Ub linearly increased (49 ± 2 ng/mL/day; r2 = 0.82, P < 0.001) 20-fold to 2170 ± 268 ng/mL on day 42. Plasma inhibited TNF-α production but stimulated IL-8 production of normal whole blood, which correlated with time-dependent Ub release (TNFα: rspearman = -0.626, P < 0.001; IL-8: rspearman = 0.427, P = 0.004). Addition of exogenous Ub (equaling day 42 concentration) to day 0-4 plasma inhibited TNF-α production by one-third of the effect detected for day 42 plasma, but also inhibited IL-8 production by 40{\%}. IL-6 production and cell proliferation was unchanged between day 0-4 plasma with or without Ub supplementation and day 42 plasma. Conclusions: Extracellular Ub release in pRBCs correlates with in vitro immunomodulatory effects and may partially contribute to transfusion-related immune modulation. Additionally, the linear kinetics of the ubiquitin release during pRBC storage suggest Ub is a suitable in vitro quality control parameter.",
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