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

Camillo Almici,Mirella Marini,Rosanna Verardi,Kurt Baeten,Cristina Skert,Domenico Russo,Giovanna Piovani,Andrea Di Palma,Gianluca Rotta,Valeria Cancelli,Simona Braga,Arabella Neva,Benedetto Bruno,Andrea Bianchetti,Paola Omedè

doi:10.1038/s41598-018-36442-9

Camillo Almici, Mirella Marini + Show 13 more

Open Access

https://doi.org/10.1038/s41598-018-36442-9

Copy DOI

Abstract

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 (ClinicalTrials.gov, 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.

Highlights

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
In order to reduce the need of invasive procedures, clinical researchers are hardly engaged in seeking specific and objective biomarkers in peripheral blood (PB) for improving GVHD diagnostic definition[6]
Network: Standardization of Circulating ENdothelIal Cells) that, utilizing a highly optimized method based on a polychromatic flowcytometry (PFC) Lyotube, proved the possibility to reach a high level standardization in Circulating Endothelial Cells (CEC) count and analysis[18,19]

Summary

Introduction

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. Allogeneic hematopoietic stem cell transplantation (allo-HSCT) has strongly improved, over the past decades, the cure rate of several onco-hematologic diseases[1] Despite of these promising results, graft-versus-host disease (GVHD) remains one of the major factors influencing patients survival[2,3]. We present the results of a comparative study of CEC counts (5 different timepoints) performed in parallel by means of an ad hoc designed PFC Lyotube and the CellSearch system in a cohort of 50 patients with malignant hematologic diseases undergoing allo-HSCT (ClinicalTrials.gov, NCT02064972). Our results can be the basis for future large-scale confirmatory study

Objectives

Methods

Results

Conclusion