Adenovirus vectors can infect mouse megakaryocytes – implications for vaccine-induced thrombosis/thrombocytopenia

Background: Adenoviruses, adenoviral vector vaccines, and gene delivery systems have been implicated in thrombosis/thrombocytopenia syndrome. The underlying cause remains unknown. We have previously hypothesized that infection of megakaryocytes (MKs), the progenitor cells of platelets, plays a centr...

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Main Authors: Simon Collett, Linda Earnest, Melissa A. Edeling, Ashley Huey Yiing Yap, Julio Carrera Montoya, Chantal Attard, Ladina Di Rago, Alison Farley, Ashley P. Ng, Paul Monagle, Joseph Torresi
Format: Article
Language:English
Published: Elsevier 2025-05-01
Series:Research and Practice in Thrombosis and Haemostasis
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Online Access:http://www.sciencedirect.com/science/article/pii/S247503792500233X
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author Simon Collett
Linda Earnest
Melissa A. Edeling
Ashley Huey Yiing Yap
Julio Carrera Montoya
Chantal Attard
Ladina Di Rago
Alison Farley
Ashley P. Ng
Paul Monagle
Joseph Torresi
author_facet Simon Collett
Linda Earnest
Melissa A. Edeling
Ashley Huey Yiing Yap
Julio Carrera Montoya
Chantal Attard
Ladina Di Rago
Alison Farley
Ashley P. Ng
Paul Monagle
Joseph Torresi
author_sort Simon Collett
collection DOAJ
description Background: Adenoviruses, adenoviral vector vaccines, and gene delivery systems have been implicated in thrombosis/thrombocytopenia syndrome. The underlying cause remains unknown. We have previously hypothesized that infection of megakaryocytes (MKs), the progenitor cells of platelets, plays a central role in adenovirus- or adenoviral vector-induced thrombosis/thrombocytopenia. Further, recent work has highlighted that the MK population comprises multiple subsets, including cells with immune function, and we propose that this MK subset is involved in adenoviral infection and the subsequent response. Objectives: To determine whether MKs are permissive to infection by recombinant adenovirus encoding the SARS-CoV-2 spike protein, and whether infection is associated with phenotypic changes. Methods: In this work, we generated adenovirus vectors based on the adenovirus 5 strain, with a green fluorescence protein reporter gene and encoding or not the SARS-CoV-2 Spike protein. Megakaryoblastic cell lines and MKs derived from mouse bone marrow and differentiated with thrombopoietin were exposed to adenoviral vectors, and infection was analyzed using flow cytometry and fluorescence microscopy. Results: The primary finding of this study is that MKs were permissive to infection by recombinant adenoviruses carrying the Spike gene of ß-SARS-CoV-2, with higher rates of infection in mature MKs. Furthermore, the effect of adenoviral infection on the cell surface proteins CD41, CD42, and CXCR4 was investigated. Cell cultures stimulated with bacterial lipopolysaccharide, with and without concurrent adenoviral infection, demonstrated that lipopolysaccharide stimulation and adenoviral infection at moderate multiplicity of infection drove increased surface expression of the α chemokine receptor CXCR4, whereas adenoviral infection at a high multiplicity of infection reduced CXCR4 expression. Conclusion: We found that adenoviral infection was higher in higher-ploidy CD41+CD42+ and CXCR4hi MKs, implicating these receptors in the MK response to adenoviral infection.
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spelling doaj-art-d0c03ea28e244e499faa4f870571ae852025-08-20T03:38:19ZengElsevierResearch and Practice in Thrombosis and Haemostasis2475-03792025-05-019410290910.1016/j.rpth.2025.102909Adenovirus vectors can infect mouse megakaryocytes – implications for vaccine-induced thrombosis/thrombocytopeniaSimon Collett0Linda Earnest1Melissa A. Edeling2Ashley Huey Yiing Yap3Julio Carrera Montoya4Chantal Attard5Ladina Di Rago6Alison Farley7Ashley P. Ng8Paul Monagle9Joseph Torresi10Department of Paediatrics, The University of Melbourne, Melbourne, Victoria, Australia; Department of Microbiology and Immunology, The Peter Doherty Institute for Infection and Immunity, The University of Melbourne, Melbourne, Victoria, Australia; Haematology Research, Murdoch Children’s Research Institute, Melbourne, Victoria, Australia; Correspondence Simon Collett, Department of Paediatrics, The University of Melbourne, Melbourne, VIC 3010, Australia.Department of Microbiology and Immunology, The Peter Doherty Institute for Infection and Immunity, The University of Melbourne, Melbourne, Victoria, AustraliaDepartment of Microbiology and Immunology, The Peter Doherty Institute for Infection and Immunity, The University of Melbourne, Melbourne, Victoria, AustraliaDepartment of Microbiology and Immunology, The Peter Doherty Institute for Infection and Immunity, The University of Melbourne, Melbourne, Victoria, AustraliaDepartment of Microbiology and Immunology, The Peter Doherty Institute for Infection and Immunity, The University of Melbourne, Melbourne, Victoria, AustraliaHaematology Research, Murdoch Children’s Research Institute, Melbourne, Victoria, AustraliaThe Walter and Eliza Hall Institute of Medical Research, Immunology, 1G Royal Parade, Melbourne, Victoria, AustraliaThe Walter and Eliza Hall Institute of Medical Research, Immunology, 1G Royal Parade, Melbourne, Victoria, AustraliaThe Walter and Eliza Hall Institute of Medical Research, Immunology, 1G Royal Parade, Melbourne, Victoria, Australia; Department of Medical Biology, The University of Melbourne, Melbourne, Victoria, Australia; Department of Clinical Haematology, Peter MacCallum Cancer Centre, The Royal Melbourne Hospital, Melbourne, Victoria, AustraliaDepartment of Paediatrics, The University of Melbourne, Melbourne, Victoria, Australia; Haematology Research, Murdoch Children’s Research Institute, Melbourne, Victoria, Australia; Department of Clinical Haematology, Royal Children’s Hospital, Melbourne, Victoria, AustraliaDepartment of Microbiology and Immunology, The Peter Doherty Institute for Infection and Immunity, The University of Melbourne, Melbourne, Victoria, AustraliaBackground: Adenoviruses, adenoviral vector vaccines, and gene delivery systems have been implicated in thrombosis/thrombocytopenia syndrome. The underlying cause remains unknown. We have previously hypothesized that infection of megakaryocytes (MKs), the progenitor cells of platelets, plays a central role in adenovirus- or adenoviral vector-induced thrombosis/thrombocytopenia. Further, recent work has highlighted that the MK population comprises multiple subsets, including cells with immune function, and we propose that this MK subset is involved in adenoviral infection and the subsequent response. Objectives: To determine whether MKs are permissive to infection by recombinant adenovirus encoding the SARS-CoV-2 spike protein, and whether infection is associated with phenotypic changes. Methods: In this work, we generated adenovirus vectors based on the adenovirus 5 strain, with a green fluorescence protein reporter gene and encoding or not the SARS-CoV-2 Spike protein. Megakaryoblastic cell lines and MKs derived from mouse bone marrow and differentiated with thrombopoietin were exposed to adenoviral vectors, and infection was analyzed using flow cytometry and fluorescence microscopy. Results: The primary finding of this study is that MKs were permissive to infection by recombinant adenoviruses carrying the Spike gene of ß-SARS-CoV-2, with higher rates of infection in mature MKs. Furthermore, the effect of adenoviral infection on the cell surface proteins CD41, CD42, and CXCR4 was investigated. Cell cultures stimulated with bacterial lipopolysaccharide, with and without concurrent adenoviral infection, demonstrated that lipopolysaccharide stimulation and adenoviral infection at moderate multiplicity of infection drove increased surface expression of the α chemokine receptor CXCR4, whereas adenoviral infection at a high multiplicity of infection reduced CXCR4 expression. Conclusion: We found that adenoviral infection was higher in higher-ploidy CD41+CD42+ and CXCR4hi MKs, implicating these receptors in the MK response to adenoviral infection.http://www.sciencedirect.com/science/article/pii/S247503792500233XAdenoviridaehemostasismegakaryocytethrombocytopeniathrombosis
spellingShingle Simon Collett
Linda Earnest
Melissa A. Edeling
Ashley Huey Yiing Yap
Julio Carrera Montoya
Chantal Attard
Ladina Di Rago
Alison Farley
Ashley P. Ng
Paul Monagle
Joseph Torresi
Adenovirus vectors can infect mouse megakaryocytes – implications for vaccine-induced thrombosis/thrombocytopenia
Research and Practice in Thrombosis and Haemostasis
Adenoviridae
hemostasis
megakaryocyte
thrombocytopenia
thrombosis
title Adenovirus vectors can infect mouse megakaryocytes – implications for vaccine-induced thrombosis/thrombocytopenia
title_full Adenovirus vectors can infect mouse megakaryocytes – implications for vaccine-induced thrombosis/thrombocytopenia
title_fullStr Adenovirus vectors can infect mouse megakaryocytes – implications for vaccine-induced thrombosis/thrombocytopenia
title_full_unstemmed Adenovirus vectors can infect mouse megakaryocytes – implications for vaccine-induced thrombosis/thrombocytopenia
title_short Adenovirus vectors can infect mouse megakaryocytes – implications for vaccine-induced thrombosis/thrombocytopenia
title_sort adenovirus vectors can infect mouse megakaryocytes implications for vaccine induced thrombosis thrombocytopenia
topic Adenoviridae
hemostasis
megakaryocyte
thrombocytopenia
thrombosis
url http://www.sciencedirect.com/science/article/pii/S247503792500233X
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