Characterization of buffy coat-derived granulocytes for clinical use: a comparison with granulocyte colony-stimulating factor/dexamethasone-pretreated donor-derived products

Vox Sang. 2017 Feb;112(2):173-182. doi: 10.1111/vox.12481. Epub 2017 Jan 25.

Abstract

Background and objectives: Buffy coat-derived granulocytes have been described as an alternative to the apheresis product from donors pretreated with dexamethasone and granulocyte colony-stimulating factor (G-CSF). The latter is - dependent on the local and national settings - obtained following a demanding and time-consuming procedure, which is undesirable in critically ill septic patients. In contrast, buffy coat-derived products have a large volume and are often heavily contaminated with red cells and platelets. We developed a new pooled buffy coat-derived product with high purity and small volume, and performed a comprehensive functional characterization of these granulocytes.

Materials and methods: We pooled ten buffy coats following the production of platelet concentrates. Saline 0·9% was added to decrease the viscosity and the product was split into plasma, red cells and a 'super' buffy coat. Functional data of the granulocytes were compared to those obtained with granulocytes from healthy controls and G-CSF/dexamethasone-pretreated donors.

Results: Buffy coat-derived granulocytes showed adhesion, chemotaxis, reactive oxygen species production, degranulation, NETosis and in vitro killing of Staphylococcus aureus, Escherichia coli and Aspergillus species comparable to control and G-CSF/dexamethasone-derived granulocytes. Candida killing was superior compared to G-CSF/dexamethasone-derived granulocytes. Immunophenotyping was normal; especially no signs of activation in the buffy coat-derived granulocytes were seen. Viability was reduced. Buffy coats are readily available in the regular blood production process and would take away the concerns around the apheresis product.

Conclusion: The product described appears a promising alternative for transfusion purposes.

Keywords: buffy coat; dexamethasone; granulocyte; granulocyte colony-stimulating factor; neutrophil; transfusion.

MeSH terms

  • Adult
  • Antigens, Surface / metabolism
  • Blood Buffy Coat / cytology*
  • Blood Component Removal
  • Blood Donors
  • Blood Platelets / cytology
  • Cell Adhesion / drug effects
  • Cell Survival / drug effects
  • Chemotaxis / drug effects
  • Dexamethasone / pharmacology*
  • Granulocyte Colony-Stimulating Factor / pharmacology*
  • Granulocytes / cytology
  • Granulocytes / drug effects*
  • Granulocytes / immunology
  • Granulocytes / metabolism
  • Humans
  • Immunophenotyping
  • Leukocyte Count
  • Male
  • NADPH Oxidases / metabolism
  • Reactive Oxygen Species / metabolism

Substances

  • Antigens, Surface
  • Reactive Oxygen Species
  • Granulocyte Colony-Stimulating Factor
  • Dexamethasone
  • NADPH Oxidases