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Chronic Lymphocytic Leukemia

International standardized approach for flow cytometric residual disease monitoring in chronic lymphocytic leukaemia

Leukemia volume 21, pages 956–964 (2007)Cite this article

Abstract

The eradication of minimal residual disease (MRD) in chronic lymphocytic leukaemia (CLL) predicts for improved outcome. However, the wide variety of MRD techniques makes it difficult to interpret and compare different clinical trials. Our aim was to develop a standardized flow cytometric CLL-MRD assay and compare it to real-time quantitative allele-specific oligonucleotide (RQ-ASO) Immunoglobulin heavy chain gene (IgH) polymerase chain reaction (PCR). Analysis of 728 paired blood and marrow samples demonstrated high concordance (87%) for patients off-therapy. Blood analysis was equally or more sensitive than marrow in 92% of samples but marrow analysis was necessary to detect MRD within 3 months of alemtuzumab therapy. Assessment of 50 CLL-specific antibody combinations identified three (CD5/CD19 with CD20/CD38, CD81/CD22 and CD79b/CD43) with low inter-laboratory variation and false-detection rates. Experienced operators demonstrated an accuracy of 95.7% (specificity 98.8%, sensitivity 91.1%) in 141 samples with 0.01–0.1% CLL. There was close correlation and 95% concordance with RQ-ASO IgH-PCR for detection of CLL above 0.01%. The proposed flow cytometry approach is applicable to all sample types and therapeutic regimes, and sufficiently rapid and sensitive to guide therapy to an MRD-negativity in real time. These techniques may be used as a tool for assessing response and comparing the efficacy of different therapeutic approaches.

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Acknowledgements

Thanks to Florence Cymbalista, Remi Letestu, Sorina Tugulea, Chris Etzler, Fiona Bennett, Ruth de Tute, Brad Dickinson and Ludmila Bezdícková for much appreciated input into the project. This work was supported by Leukaemia Research Fund, UK; AIRC Milano, Italy; MIUR, Italy; National Cancer Institute Grant R21 CA101332 and U10 CA101140, US; Ben Eckenhoff fund, US; Instituto de Salud Carlos III, FIS 04/1051, Spain.

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Authors and Affiliations

  1. HMDS, Leeds Teaching Hospitals, Leeds, UK

    A C Rawstron, P A S Evans & P Hillmen

  2. Departments of Hematology and Pathology, Unitat d’Hematopatologia, Hospital Clínic, Institut d’Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain

    N Villamor, D Colomer, C Moreno & E Montserrat

  3. Departments of Hematology and Pathology, Unitat d’Hematopathologia, Hospital Clínic, Institut d’Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain

    N Villamor, D Colomer, C Moreno & E Montserrat

  4. Second Department of Medicine, University of Schleswig-Holstein, Campus Kiel, Germany

    M Ritgen, S Böttcher & M Kneba

  5. Department of Oncology, Università Vita-Salute San Raffaele and Istituto Scientifico San Raffaele, Milano, Italy

    P Ghia & F Caligaris-Cappio

  6. Department of Medicine, Division of Hematology, Stanford University School of Medicine, Stanford, CA, USA

    J L Zehnder, Y Natkunam & S E Coutre

  7. Division of Hematology and Oncology, The Arthur James Comprehensive Cancer Center, The Ohio State University, Columbus, OH, USA

    G Lozanski & J C Byrd

  8. Laboratory of Cytometry, Institute for Cancer Research and Treatment (IRCC), Candiolo, Italy

    M Geuna

  9. Department of Medicine, Division of Hematology/Oncology, University of California, San Diego, UK

    E D Avery, L Z Rassenti & T J Kipps

  10. Klinik I für Innere Medizin, Universität zu Köln, Köln, Germany

    M J Hallek

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  1. A C Rawstron
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  2. N Villamor
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  3. M Ritgen
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  4. S Böttcher
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  5. P Ghia
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  6. J L Zehnder
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  7. G Lozanski
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  8. D Colomer
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  9. C Moreno
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  10. M Geuna
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  11. P A S Evans
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  12. Y Natkunam
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  13. S E Coutre
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  14. E D Avery
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  15. L Z Rassenti
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  16. T J Kipps
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  17. F Caligaris-Cappio
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  18. M Kneba
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  19. J C Byrd
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  20. M J Hallek
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  21. E Montserrat
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  22. P Hillmen
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Corresponding author

Correspondence to A C Rawstron.

Additional information

On behalf of European Research Initiative on CLL (www.ericll.org) and the CLL Research Consortium (http://cll.ucsd.edu/)

Supplementary Information accompanies the paper on the Leukemia website (http://www.nature.com/leu)

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Rawstron, A., Villamor, N., Ritgen, M. et al. International standardized approach for flow cytometric residual disease monitoring in chronic lymphocytic leukaemia. Leukemia 21, 956–964 (2007). https://doi.org/10.1038/sj.leu.2404584

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