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Peer-reviewedCC BY 4.0Frontiers in Immunology

Extracellular vesicle phenotypic patterns associated with CMV, EBV and HHV-6 reactivation after haploidentical transplantation with post-transplant cyclophosphamide

  • Giuseppe Lia1,2
  • Clara Di Vito3,4
  • Stefania Bruno5
  • Jacopo Mariotti6
  • Alessia Melis2
  • Michela Calvi3,4
  • Luisa Giaccone1,2
  • Armando Santoro6
  • Stefania Bramanti6
  • Luca Castagna6
  • Andrea Evangelista7
  • Domenico Mavilio3,4
1 more authors
  • Benedetto Bruno1,2
7 affiliations
  1. 1Division of Hematology, Department of Oncology, A.O.U. Città della Salute e della Scienza di Torino
  2. 2Department of Molecular Biotechnology and Health Sciences, University of Torino
  3. 3Department of Medical Biotechnologies and Translational Medicine (BioMeTra), University of Milan
  4. 4Unit of Clinical and Experimental Immunology, Istituti di Ricovero e Cura a Carattere Scientifico (IRCCS) Humanitas Research Hospital, Rozzano
  5. 5Department of Medical Sciences, University of Torino
  6. 6Bone Marrow Transplant Unit, IRCCS Humanitas Research Hospital
  7. 7Clinical Epidemiology, A.O.U. Città della Salute e della Scienza di Torino
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Abstract

Introduction: Human cytomegalovirus (CMV), Epstein–Barr virus (EBV), and human herpesvirus 6 (HHV-6) remain important causes of morbidity after allogeneic hematopoietic cell transplantation (HCT), yet biomarkers predicting viral reactivation are lacking. Extracellular vesicles (EVs) have emerged as promising liquid-biopsy tools in inflammatory and infectious diseases.

Methods: In this exploratory biomarker discovery study, 32 consecutive recipients of haploidentical HCT with post-transplant cyclophosphamide were prospectively enrolled. EV surface marker profiles were assessed by flow cytometry before transplantation and at predefined time points during the first three months post-HCT. Associations between EV phenotypes and viral reactivation were evaluated using logistic regression and time-varying Cox models.

Results: CMV, EBV, and HHV-6 reactivation occurred in 20, 11, and 17 patients, respectively. Multivariable analyses showed that different EV phenotypic patterns were associated with CMV, EBV, and HHV-6 reactivation. CMV reactivation was associated with increased EV expression of CD44, CD86, CD140a, and CD144. EBV reactivation correlated with reduced CD30-positive EV concentrations and increased KRT18 expression. HHV-6 reactivation was associated with altered CD26, CD86, CD120a, and CD144 EV profiles. These associations remained significant after adjustment for acute graft-versus-host disease and concomitant viral infections.

Discussion: Collectively, these exploratory findings support further validation of circulating EV phenotypes as candidate biomarkers for predicting and monitoring viral reactivation after haploidentical HCT.

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Cite

BibTeXRIS
BibTeX
@article{lia2026extracellular,
  title = {{Extracellular vesicle phenotypic patterns associated with CMV, EBV and HHV-6 reactivation after haploidentical transplantation with post-transplant cyclophosphamide}},
  author = {Giuseppe Lia and Clara Di Vito and Stefania Bruno and Jacopo Mariotti and Alessia Melis and Michela Calvi and Luisa Giaccone and Armando Santoro and Stefania Bramanti and Luca Castagna and Andrea Evangelista and Domenico Mavilio and Benedetto Bruno},
  journal = {Frontiers in Immunology},
  year = {2026},
  month = oct,
  doi = {10.3389/fimmu.2026.1937675},
  url = {https://www.frontiersin.org/articles/10.3389/fimmu.2026.1937675/full},
  note = {CC BY 4.0},
}
RIS
TY  - JOUR
TI  - Extracellular vesicle phenotypic patterns associated with CMV, EBV and HHV-6 reactivation after haploidentical transplantation with post-transplant cyclophosphamide
AU  - Giuseppe Lia
AU  - Clara Di Vito
AU  - Stefania Bruno
AU  - Jacopo Mariotti
AU  - Alessia Melis
AU  - Michela Calvi
AU  - Luisa Giaccone
AU  - Armando Santoro
AU  - Stefania Bramanti
AU  - Luca Castagna
AU  - Andrea Evangelista
AU  - Domenico Mavilio
AU  - Benedetto Bruno
PY  - 2026
DA  - 2026/10/05
JO  - Frontiers in Immunology
DO  - 10.3389/fimmu.2026.1937675
UR  - https://www.frontiersin.org/articles/10.3389/fimmu.2026.1937675/full
AB  - Introduction: Human cytomegalovirus (CMV), Epstein–Barr virus (EBV), and human herpesvirus 6 (HHV-6) remain important causes of morbidity after allogeneic hematopoietic cell transplantation (HCT), yet biomarkers predicting viral reactivation are lacking. Extracellular vesicles (EVs) have emerged as promising liquid-biopsy tools in inflammatory and infectious diseases.  Methods: In this exploratory biomarker discovery study, 32 consecutive recipients of haploidentical HCT with post-transplant cyclophosphamide were prospectively enrolled. EV surface marker profiles were assessed by flow cytometry before transplantation and at predefined time points during the first three months post-HCT. Associations between EV phenotypes and viral reactivation were evaluated using logistic regression and time-varying Cox models.  Results: CMV, EBV, and HHV-6 reactivation occurred in 20, 11, and 17 patients, respectively. Multivariable analyses showed that different EV phenotypic patterns were associated with CMV, EBV, and HHV-6 reactivation. CMV reactivation was associated with increased EV expression of CD44, CD86, CD140a, and CD144. EBV reactivation correlated with reduced CD30-positive EV concentrations and increased KRT18 expression. HHV-6 reactivation was associated with altered CD26, CD86, CD120a, and CD144 EV profiles. These associations remained significant after adjustment for acute graft-versus-host disease and concomitant viral infections.  Discussion: Collectively, these exploratory findings support further validation of circulating EV phenotypes as candidate biomarkers for predicting and monitoring viral reactivation after haploidentical HCT.
SN  - 1664-3224
ER  -