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

  • Shimrit Ringelstein-Harlev
  • Irit Avivi
  • Mona Fanadka
  • Netanel A. Horowitz
  • Tami Katz
Original Article

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.

Keywords

Chronic lymphocytic leukemia (CLL) Regulatory B-cells (Bregs) Interleukin 10 (IL-10) Toll-like receptor 9 (TLR9) 

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

Notes

Acknowledgements

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

Compliance with ethical standards

Conflict of interest

There are no conflicts to declare.

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Shimrit Ringelstein-Harlev
    • 1
  • Irit Avivi
    • 3
    • 4
  • Mona Fanadka
    • 2
  • Netanel A. Horowitz
    • 1
    • 2
  • Tami Katz
    • 1
    • 2
  1. 1.Department of Hematology and Bone Marrow TransplantationRambam Health Care CampusHaifaIsrael
  2. 2.Bruce Rappaport Faculty of MedicineTechnion, Israel Institute of TechnologyHaifaIsrael
  3. 3.Department of Hematology and Bone Marrow TransplantationTel Aviv Sourasky Medical CenterTel AvivIsrael
  4. 4.Sackler School of MedicineTel Aviv UniversityTel AvivIsrael

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