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Chronic Lymphocytic Leukemia

Analysis of CD16+CD56dim NK cells from CLL patients: evidence supporting a therapeutic strategy with optimized anti-CD20 monoclonal antibodies

Abstract

Although anti-CD20 monoclonal antibodies (mAbs) show promise for the treatment of chronic lymphocytic leukemia (CLL), the success of the anti-CD20 mAb rituximab in CLL treatment has been limited. Novel anti-CD20 mAbs with more potent cytotoxic activity have recently been engineered, but so far most have only been tested in vitro with natural killer (NK) cells from healthy donors. Because it is still unclear whether these optimized cytotoxic mAbs will improve NK-cell killing of tumor cells in CLL patients, we characterized the relevant phenotypic and functional features of NK cells from CLL patients in detail. Expression of inhibitory and activating NK-cell receptors and of Fc gamma receptor IIIA (FcγRIIIA) is well preserved in CD16+CD56dim cytotoxic NK cells from these patients, independently of disease progression. These cells are fully functional following cytokine stimulation. In addition, the FcγRIIIA-optimized LFB-R603 anti-CD20 mAb mediates 100 times greater antibody-dependent cell-mediated cytotoxicity by NK cells from CLL patients and healthy donors than rituximab. Enhanced degranulation against autologous B-CLL cells is observed at lower concentrations of LFB-R603 than rituximab, regardless of CLL prognostic factors. These findings strongly justify further clinical development of anti-CD20 mAbs optimized for FcγR engagement in CLL patients.

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Acknowledgements

We thank Z Azgui, K Maloum, F Nguyen-Khac, JL Binet, N Marchay, H M’Kada and S Nguyen-Quoc, from the Hematology Department at Pitié-Salpêtrière Hospital (AP-HP, Paris, France) for the recruitment of CLL patients. We thank F Gandjbakhch, C Poulain and the personnel from the Department of Rheumatology at Pitié-Salpêtrière Hospital (AP-HP, Paris, France) and V Siguret, E Pautas and the personnel from the Gerontology Department at Charles-Foix Hospital (AP-HP, Ivry, France) for the recruitment of the control group. P Bonnemye, M Brissard, S Gueguen, M Boudjoghra and A Grelier are acknowledged for their technical assistance. This study was supported in part by funds from the Laboratoire Français de Fractionnement et des Biotechnologies (LFB, Les Ulis, France), the association la Ligue contre le cancer (RS08/75-4) and INSERM. The anti-CD20 LFB-R603 antibody was purchased by LFB.

Author contributions

MLT, CAD, JLT, HMB and VV conceived and designed the study; MLT and JD performed the flow cytometric and functional analyses; CP performed the flow cytometric selection; EC provided cytogenetic analyses; FD provided IgHV sequences; HMB cared for the involved patients, provided clinical and diagnosis/prognosis cytometric data; MLT, JD, CdR, CAD, JLT, HMB and VV analyzed the flow cytometric and functional data; MLT, JLT, HMB and VV wrote the manuscript; JD, CdR, CP, CAD, EC, FD, PD and JFP critically read and approved the manuscript.

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Correspondence to M Le Garff-Tavernier.

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Competing interests

J Decocq, C de Romeuf and JF Prost are employed by LFB, whose potential product was studied in the present report. CA Dutertre was supported by a CIFRE fellowship from the ANRT and LFB (#134/2004) between 2004 and 2007. J-L Teillaud was a LFB consultant until June 2007, which covers part of the study period. Three of the authors, C de Romeuf, JF Prost and JL Teillaud, are designated as inventors on patent application WO2006064121 owned by LFB Biotechnologies, and claim specific therapeutic use of the antibody studied in this report. The other authors declare no conflict of interest.

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Le Garff-Tavernier, M., Decocq, J., de Romeuf, C. et al. Analysis of CD16+CD56dim NK cells from CLL patients: evidence supporting a therapeutic strategy with optimized anti-CD20 monoclonal antibodies. Leukemia 25, 101–109 (2011). https://doi.org/10.1038/leu.2010.240

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Keywords

  • chronic lymphocytic leukemia (CLL)
  • natural killer (NK) cells
  • anti-CD20
  • monoclonal antibodies

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