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Sci Rep. 2019 Jan 14;9(1):87. doi: 10.1038/s41598-018-36442-9.

Counting circulating endothelial cells in allo-HSCT: an ad hoc designed polychromatic flowcytometry-based panel versus the CellSearch System.

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Laboratory for Stem Cells Manipulation and Cryopreservation, Dpt of Transfusion Medicine, ASST Spedali Civili, Brescia, Italy.
Laboratory for Stem Cells Manipulation and Cryopreservation, Dpt of Transfusion Medicine, ASST Spedali Civili, Brescia, Italy.
Chair of Hematology, Unit of Blood Diseases and Stem Cell Transplantation, University of Brescia, ASST Spedali Civili, Brescia, Italy.
BMT Unit, Department of Molecular Biotechnology and Health Sciences, University of Torino, A.O.U Città della Salute e della Scienza di Torino, Torino, Italy.
Biology and Genetics Division, Department of Molecular and Translational Medicine, University of Brescia, Brescia, Italy.
Global Scientific and Medical Affairs, Janssen Diagnostics, Beerse, Belgium.
Scientific Affairs, BD Biosciences Italia, Milano, Italy.


Physio-pathologic interrelationships between endothelial layer and graft-versus-host disease (GVHD) have been described leading to assess the entity "endothelial GVHD" as the early step for clinical manifestations of acute GVHD. The availability of the CellSearch system has allowed us to monitor Circulating Endothelial Cells (CEC) changes in allogeneic hematopoietic stem cell transplantation (allo-HSCT) as useful tool to help clinicians in GVHD diagnostic definition. We have compared CEC counts generated by an ad hoc designed polychromatic-flowcytometry (PFC) Lyotube with those of the CellSearch system. CEC were counted in parallel at 5 timepoints in 50 patients with malignant hematologic disorders undergoing allo-HSCT (, NCT02064972). Spearman rank correlation showed significant association between CEC values at all time points (p = 0.0001). The limits of agreement was demonstrated by Bland Altman plot analysis, showing bias not significant at T1, T3, T4, while at T2 and T5 resulted not estimable. Moreover, Passing Bablok regression analysis showed not significant differences between BD Lyotube and CellSearch system. We show that CEC counts, generated with either the CellSearch system or the PFC-based panel, have a superimposable kinetic in allo-HSCT patients and that both counting procedures hold the potential to enter clinical routine as a suitable tool to assist clinicians in GVHD diagnosis.

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