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

  • Richard Charles Duggleby
  • J. Alejandro Madrigal
Part of the Methods in Molecular Biology book series (MIMB, volume 1109)


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.

Key words

Hematopoietic stem cell therapy Cellular reconstitution B cells NK cells T cells Regulatory T cells Dendritic cells Whole blood flow cytometry 



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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Copyright information

© Springer Science+Business Media, New York 2014

Authors and Affiliations

  • Richard Charles Duggleby
    • 1
  • J. Alejandro Madrigal
    • 2
  1. 1.Anthony Nolan Research InstituteRoyal Free HospitalLondonUK
  2. 2.Anthony Nolan Research InstituteUniversity College LondonLondonUK

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