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Chronic lymphocytic leukemia cells acquire regulatory B-cell properties in response to TLR9 and CD40 activation

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Abstract

Circulating chronic lymphocytic leukemia (CLL) cells share phenotypic features with certain subsets of regulatory B-cells (Bregs). The latter cells have been reported to negatively regulate immune cell responses, mostly by provision of IL-10. The purpose of the current study was to identify and delineate Breg properties of CLL cells. B-cells and T-cells were obtained from the peripheral blood of untreated CLL patients diagnosed according to the 2008 Guidelines of the International Workshop on Chronic Lymphocytic Leukemia. Co-culture assays were used to examine the ability of CLL cells to suppress autologous T-cell immune responses. IL-10 potency of CLL cells was assessed following stimulation with activators of the toll-like receptor 9 (TLR9) or CD40 and was correlated with the inhibitory activity of the cells. TLR9-activated CLL cells were found to increase the frequency of CD4+CD25hiFOXp3+ regulatory T-cells (Tregs) and to inhibit autologous CD4+ T-cell proliferation. This signaling cascade proved to control IL-10 generation in CLL cells, which in turn promoted the inhibition of T-cell proliferation by CLL cells. However, CD40 activation of CLL cells, while exhibiting a similar ability to augment Treg frequency, did not either affect IL-10 generation or T-cell proliferation. In conclusion, CLL cells demonstrate a unique clonal quality of adopting Breg properties which promote modulation of T-cell characteristics. TLR9 appears to be a potent activator of regulatory abilities in CLL cells, possibly contributing to preferential immune escape of TLR9-responsive cells.

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Abbreviations

ASH:

American Society of Hematology

APC:

Allophycocyanin

BCR:

B-cell receptor

Bregs:

Regulatory B-cells

BV:

Brilliant violet

CD40L:

CD40 ligand

CFSE:

Carboxyfluorescein diacetate succinimidyl ester

CLL:

Chronic lymphocytic leukemia

FISH:

Fluorescence in situ hybridization

IgVH:

Immunoglobulin variable region heavy chain

IRAK4:

Interleukin-1 receptor-associated kinase 4

ODN:

Oligodeoxynucleotide

qPCR:

Quantitative polymerase chain reaction

siRNA:

Small interfering RNA

Tregs:

Regulatory T-cells

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Acknowledgements

The authors wish to acknowledge with thanks the assistance of Sonia Kamenetsky in the preparation of the manuscript.

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Author notes

  1. Shimrit Ringelstein-Harlev and Irit Avivi have equally contributed.

Authors and Affiliations

  1. Department of Hematology and Bone Marrow Transplantation, Rambam Health Care Campus, 8, Ha’Aliya Street, 3109601, Haifa, Israel

    Shimrit Ringelstein-Harlev, Netanel A. Horowitz & Tami Katz

  2. Bruce Rappaport Faculty of Medicine, Technion, Israel Institute of Technology, Haifa, Israel

    Mona Fanadka, Netanel A. Horowitz & Tami Katz

  3. Department of Hematology and Bone Marrow Transplantation, Tel Aviv Sourasky Medical Center, Tel Aviv, Israel

    Irit Avivi

  4. Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel

    Irit Avivi

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  1. Shimrit Ringelstein-Harlev
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Correspondence to Shimrit Ringelstein-Harlev.

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Ringelstein-Harlev, S., Avivi, I., Fanadka, M. et al. Chronic lymphocytic leukemia cells acquire regulatory B-cell properties in response to TLR9 and CD40 activation. Cancer Immunol Immunother 67, 739–748 (2018). https://doi.org/10.1007/s00262-018-2128-x

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  • DOI: https://doi.org/10.1007/s00262-018-2128-x

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