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CHRONIC LYMPHOCYTIC LEUKEMIA

Activation and expansion of T-follicular helper cells in chronic lymphocytic leukemia nurselike cell co-cultures

Leukemia volume 36, pages 1324–1335 (2022)Cite this article

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Abstract

Interactions between chronic lymphocytic leukemia (CLL) cells and T-cell subsets in the lymph node microenvironment are thought to play a central role in disease biology. To study these interactions in a model of the CLL lymph node microenvironment, we characterized T-cell subsets in CLL nurselike cell (NLC) co-cultures. We focused on T-follicular helper (Tfh) cells, which are characterized by CXCR5 expression and localization to B-cell follicles. In co-cultures from 28 different CLL patients, we detected an expansion of Tfh cells based on PD-1, BCL6, and ICOS expression, with increased IL-21 and downmodulated CD40L surface expression. Regulatory T cells (Treg), which promote immune tolerance, also expanded in NLC co-cultures. T-cell receptor (TR) gene repertoire analyses confirmed the clonal expansion of CD4+ T cells, with an enrichment of TR clonotypes commonly expanded also in primary CLL samples. Multicolor confocal microscopy revealed that Tfh, but not Treg co-localize with proliferating CLL cells in CLL lymph node sections. Collectively, these data provide new insight into the cellular and molecular cross-talk between CLL and T-cell subsets, resulting in clonal expansion of T-helper cells and interaction of Tfh cells with proliferating CLL cells which may open new avenues for therapeutic targeting.

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Fig. 1: T cell subsets in NLC co-cultures.
Fig. 2: Tfh cells acquire a germinal and memory phenotype in NLC co-culture.
Fig. 3: Germinal and memory Tfh cells show expression of BCL6 and ICOS.
Fig. 4: T cells in NLC co-cultures modulate CD40L and acquire markers of activation and proliferation.
Fig. 5: TR gene repertoire changes associated with clonal expansion of CD4+ T-helper cells in NLC co-cultures.
Fig. 6: Close physical association between proliferative CLL and Tfh cells in LN tissue.
Fig. 7: Tregs in LN tissue are in close spatial proximity to others CD4+ cells.

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Acknowledgements

The authors are grateful to all the patients who provided samples for this study. They also thank Kostas Stamatopoulos for contributing data from the TR clonotype database held at the Institute of Applied Biosciences at CERTH and for critical reading the paper. The authors thank the Nikon Imaging Facility at King’s College London. The Advanced Cytometry & Sorting Core Facility at MD Anderson is supported by NCI P30CA016672. PEMP is supported by MRC Grant MR/T005106/1. This work was supported by the ERA-NET on Translational Cancer Research (TRANSCAN-2) project code 5041673 (EV) and the generous philanthropic contributions to The University of Texas MD Anderson Cancer Center Moon Shots ProgramTM (JAB), and in part by CLL Global Research Foundation grants (JAB).

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Authors and Affiliations

  1. Department of Leukemia, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    Alicia M. Vaca, Mariela Sivina, Ekaterina Kim, Alessandra Ferrajoli, Zeev Estrov, William G. Wierda & Jan A. Burger

  2. School of Cancer and Pharmaceutical Sciences, Faculty of Life Sciences & Medicine, King’s College London, London, UK

    Nikolaos Ioannou, Piers E. M. Patten & Alan G. Ramsay

  3. Institute of Applied Biosciences, Center for Research and Technology Hellas, Thessaloniki, Greece

    Elisavet Vlachonikola

  4. Department of Stem Cell Transplantation and Hematopoietic Biology & Malignancy, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    Karen Clise-Dwyer, Dan Li & Qing Ma

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  1. Alicia M. Vaca
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Contributions

AMV performed NLC co-cultures, flow cytometry, isolation of lymphocyte populations, DNA extraction, TCR repertoire analyses, collected clinical information, analyzed the data and results, designed the figures, and wrote the paper with JAB; NI performed microscopy and analyzed the data with AR; MS assisted with data interpretation and reviewed the manuscript. EV assisted with TCR repertoire data interpretation. KCD contributed to study design. EK contributed with sample collection, storage, and reviewed the paper. DL and QM contributed to study design. AF, ZE, and WGW contributed to clinical patient management. PP contributed with CLL LNs sample collection. JAB designed and supervised the study. All authors reviewed the paper and approved the final version.

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Correspondence to Jan A. Burger.

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Vaca, A.M., Ioannou, N., Sivina, M. et al. Activation and expansion of T-follicular helper cells in chronic lymphocytic leukemia nurselike cell co-cultures. Leukemia 36, 1324–1335 (2022). https://doi.org/10.1038/s41375-022-01519-y

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