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Methods of Detection of Immune Reconstitution and T Regulatory Cells by Flow Cytometry

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Bone Marrow and Stem Cell Transplantation

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1109))

Abstract

Allogeneic hematopoietic stem cell therapy (HSCT) remains one of the few curative treatments for high-risk hematological malignancies (high-risk leukemia, myelodysplastic syndromes, advanced myeloproliferative disorders, high-risk lymphomas, and multiple myeloma) and is currently applied in more than 15,000 patients per year in Europe. Following HSCT, patients experience a period of reconstitution of the immune system, which seems to be highly dependent on conditioning, immunosuppression regimes, and the level of adverse events the patients experience. During this reconstitution period, the patient is immune compromised and susceptible to opportunistic infections and disease relapse. Consequently, a large number of clinical studies have been devoted to monitoring the recovery of the immune system following HSCT in the hopes of determining which cellular subsets are indicative of a favorable outcome. In this chapter we review the methods that have been employed to monitor the immune reconstitution and what clinical observations have been made. Of particular interest is the regulatory T cell (Treg) subset, which has been associated with tolerance and has been the subject of recent clinical trials as a possible cellular therapy for rejection reactions. Finally we will detail a proposed methodology for the flow cytometric assessment of cellular reconstitution post-HSCT.

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Acknowledgements

The authors would like to thank Dr Bronwen Shaw, Dr Sameer Tulpule, Damini Tewari, and Vikesh Devlia, Anthony Nolan Research Institute, London, UK, and Dr John Girdlestone and Dr Cristina Navarrete, H&I Department, NHS Blood and Transplant, Colindale, UK, for developing the panel on which Table 1 is based. The authors would also like to thank Dr Sergio Querol, Banc de Sang i Teixits, Barcelona, Spain, and medical consultant for the Anthony Nolan Cell Therapy Centre, Nottingham Trent University, Nottingham, UK, for his assistance in compiling this chapter.

Disclosure The authors certify that they have no affiliation with or financial involvement in any organization or entity with a direct financial interest in the subject matter or materials discussed in this manuscript.

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  1. Anthony Nolan Research Institute, Royal Free Hospital, London, UK

    Richard Charles Duggleby

  2. Anthony Nolan Research Institute, University College London, London, UK

    J. Alejandro Madrigal

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  2. J. Alejandro Madrigal
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Editors and Affiliations

  1. Department of Hematology, Ankara University, Ankara, Turkey

    Meral Beksaç

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© 2014 Springer Science+Business Media, New York

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Duggleby, R.C., Madrigal, J.A. (2014). Methods of Detection of Immune Reconstitution and T Regulatory Cells by Flow Cytometry. In: Beksaç, M. (eds) Bone Marrow and Stem Cell Transplantation. Methods in Molecular Biology, vol 1109. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4614-9437-9_10

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  • DOI: https://doi.org/10.1007/978-1-4614-9437-9_10

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4614-9436-2

  • Online ISBN: 978-1-4614-9437-9

  • eBook Packages: Springer Protocols

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