Chronic lymphocytic leukemia

CXCL13 plasma levels function as a biomarker for disease activity in patients with chronic lymphocytic leukemia


The chemoattractant CXCL13 organizes the cellular architecture of B-cell follicles and germinal centers. During adaptive immune responses, CXCL13 plasma concentrations transiently increase and function as a biomarker for normal germinal center activity. Chronic lymphocytic leukemia (CLL) cells express high levels of CXCR5, the receptor for CXCL13, and proliferate in pseudofollicles within secondary lymphoid organs (SLO). Given the morphologic and functional similarities between normal and CLL B-cell expansion in SLO, we hypothesized that CXCL13 plasma concentrations would correlate with CLL disease activity and progression. We analyzed CXCL13 plasma concentrations in 400 CLL patients and correlated the findings with other prognostic markers, time to treatment (TTT), CCL3 and CCL4 plasma concentrations, and in vivo CLL cell proliferation. We found that CXCL13 plasma concentrations were higher in CLL patients with active and advanced stage disease, resulting in a significantly shorter TTT. Accordingly, high CXCL13 levels correlated with other markers of disease activity and CCL3 levels. Higher CLL cell birth rates in vivo also associated with higher CXCL13 plasma concentrations. Interestingly, elevated CXCL13 plasma levels normalized during ibrutinib therapy, and increased in ibrutinib resistance patients. Collectively, these studies emphasize the importance of CXCL13 in crosstalk between CLL cells and the SLO microenvironment.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Fig. 1: CXCL13 plasma concentrations in CLL patients with lower- or higher-risk disease features.
Fig. 2: Distribution of CXCL13 plasma levels and probability of treatment free survival.
Fig. 3: CXCL13 plasma concentrations in CLL patients treated with ibrutinib.
Fig. 4: Correlation between SPD in the lymph nodes, percentage of CLL in bone marrow and in vivo CLL cell birth rate.


  1. 1.

    Burger JA, Wiestner A. Targeting B cell receptor signalling in cancer: preclinical and clinical advances. Nat Rev. 2018;18:148–67.

    CAS  Article  Google Scholar 

  2. 2.

    Stein H, Bonk A, Tolksdorf G, Lennert K, Rodt H, Gerdes J. Immunohistologic analysis of the organization of normal lymphoid tissue and non-Hodgkin’s lymphomas. J Histochem Cytochem. 1980;28:746–60.

    CAS  Article  Google Scholar 

  3. 3.

    Patten PE, Buggins AG, Richards J, Wotherspoon A, Salisbury J, Mufti GJ, et al. CD38 expression in chronic lymphocytic leukemia is regulated by the tumor microenvironment. Blood. 2008;111:5173–81.

    CAS  Article  Google Scholar 

  4. 4.

    Herishanu Y, Perez-Galan P, Liu D, Biancotto A, Pittaluga S, Vire B, et al. The lymph node microenvironment promotes B-cell receptor signaling, NF-kappaB activation, and tumor proliferation in chronic lymphocytic leukemia. Blood. 2011;117:563–74.

    CAS  Article  Google Scholar 

  5. 5.

    Bajenoff M, Egen JG, Koo LY, Laugier JP, Brau F, Glaichenhaus N, et al. Stromal cell networks regulate lymphocyte entry, migration, and territoriality in lymph nodes. Immunity. 2006;25:989–1001.

    CAS  Article  Google Scholar 

  6. 6.

    Allen CD, Ansel KM, Low C, Lesley R, Tamamura H, Fujii N, et al. Germinal center dark and light zone organization is mediated by CXCR4 and CXCR5. Nat Immunol. 2004;5:943–52.

    CAS  Article  Google Scholar 

  7. 7.

    Schwickert TA, Lindquist RL, Shakhar G, Livshits G, Skokos D, Kosco-Vilbois MH, et al. In vivo imaging of germinal centres reveals a dynamic open structure. Nature. 2007;446:83–7.

    CAS  Article  Google Scholar 

  8. 8.

    Allen CD, Okada T, Tang HL, Cyster JG. Imaging of germinal center selection events during affinity maturation. Science. 2007;315:528–31.

    CAS  Article  Google Scholar 

  9. 9.

    Ansel KM, Ngo VN, Hyman PL, Luther SA, Forster R, Sedgwick JD, et al. A chemokine-driven positive feedback loop organizes lymphoid follicles. Nature. 2000;406:309–14.

    CAS  Article  Google Scholar 

  10. 10.

    Victora GD, Nussenzweig MC. Germinal centers. Annu Rev Immunol. 2012;30:429–57.

    CAS  Article  Google Scholar 

  11. 11.

    Cinamon G, Zachariah MA, Lam OM, Foss FW Jr, Cyster JG. Follicular shuttling of marginal zone B cells facilitates antigen transport. Nat Immunol. 2008;9:54–62.

    CAS  Article  Google Scholar 

  12. 12.

    Mueller SN, Germain RN. Stromal cell contributions to the homeostasis and functionality of the immune system. Nat Rev Immunol. 2009;9:618–29.

    CAS  Article  Google Scholar 

  13. 13.

    Saez de Guinoa J, Barrio L, Mellado M, Carrasco YR. CXCL13/CXCR5 signaling enhances BCR-triggered B-cell activation by shaping cell dynamics. Blood. 2011;118:1560–9.

    CAS  Article  Google Scholar 

  14. 14.

    Havenar-Daughton C, Lindqvist M, Heit A, Wu JE, Reiss SM, Kendric K, et al. CXCL13 is a plasma biomarker of germinal center activity. Proc Natl Acad Sci USA. 2016;113:2702–7.

    CAS  Article  Google Scholar 

  15. 15.

    Mehraj V, Ramendra R, Isnard S, Dupuy FP, Lebouche B, Costiniuk C, et al. CXCL13 as a biomarker of immune activation during early and chronic HIV infection. Front Immunol. 2019;10:289.

    CAS  Article  Google Scholar 

  16. 16.

    Burkle A, Niedermeier M, Schmitt-Graff A, Wierda WG, Keating MJ, Burger JA. Overexpression of the CXCR5 chemokine receptor, and its ligand, CXCL13 in B-cell chronic lymphocytic leukemia. Blood. 2007;110:3316–25.

    Article  Google Scholar 

  17. 17.

    Durig J, Schmucker U, Duhrsen U. Differential expression of chemokine receptors in B cell malignancies. Leukemia. 2001;15:752–6.

    CAS  Article  Google Scholar 

  18. 18.

    Lopez-Giral S, Quintana NE, Cabrerizo M, Alfonso-Perez M, Sala-Valdes M, De Soria VG, et al. Chemokine receptors that mediate B cell homing to secondary lymphoid tissues are highly expressed in B cell chronic lymphocytic leukemia and non-Hodgkin lymphomas with widespread nodular dissemination. J Leukoc Biol. 2004;76:462–71.

    CAS  Article  Google Scholar 

  19. 19.

    Trentin L, Cabrelle A, Facco M, Carollo D, Miorin M, Tosoni A, et al. Homeostatic chemokines drive migration of malignant B cells in patients with non-Hodgkin lymphomas. Blood. 2004;104:502–8.

    CAS  Article  Google Scholar 

  20. 20.

    Stache V, Verlaat L, Gatjen M, Heinig K, Westermann J, Rehm A, et al. The splenic marginal zone shapes the phenotype of leukemia B cells and facilitates their niche-specific retention and survival. Oncoimmunology. 2017;6:e1323155.

    Article  Google Scholar 

  21. 21.

    Burger JA, Quiroga MP, Hartmann E, Burkle A, Wierda WG, Keating MJ, et al. High-level expression of the T-cell chemokines CCL3 and CCL4 by chronic lymphocytic leukemia B cells in nurselike cell cocultures and after BCR stimulation. Blood. 2009;113:3050–8.

    CAS  Article  Google Scholar 

  22. 22.

    Hartmann EM, Rudelius M, Burger JA, Rosenwald A. CCL3 chemokine expression by chronic lymphocytic leukemia cells orchestrates the composition of the microenvironment in lymph node infiltrates. Leuk Lymphoma. 2016;57:563–71.

    CAS  Article  Google Scholar 

  23. 23.

    Benet ZL, Marthi M, Ke F, Wu R, Turner JS, Gabayre JB, et al. CCL3 promotes germinal center B cells sampling by follicular regulatory T cells in murine lymph nodes. Front Immunol. 2018;9:2044.

    Article  Google Scholar 

  24. 24.

    Hallek M, Cheson BD, Catovsky D, Caligaris-Cappio F, Dighiero G, Dohner H, et al. Guidelines for the diagnosis and treatment of chronic lymphocytic leukemia: a report from the International Workshop on Chronic Lymphocytic Leukemia updating the National Cancer Institute-Working Group 1996 guidelines. Blood. 2008;111:5446–56.

    CAS  Article  Google Scholar 

  25. 25.

    Burger JA, Li KW, Keating MJ, Sivina M, Amer AM, Garg N, et al. Leukemia cell proliferation and death in chronic lymphocytic leukemia patients on therapy with the BTK inhibitor ibrutinib. JCI insight. 2017;2:e89904.

    Article  Google Scholar 

  26. 26.

    Luther SA, Lopez T, Bai W, Hanahan D, Cyster JG. BLC expression in pancreatic islets causes B cell recruitment and lymphotoxin-dependent lymphoid neogenesis. Immunity. 2000;12:471–81.

    CAS  Article  Google Scholar 

  27. 27.

    Lee HT, Shiao YM, Wu TH, Chen WS, Hsu YH, Tsai SF, et al. Serum BLC/CXCL13 concentrations and renal expression of CXCL13/CXCR5 in patients with systemic lupus erythematosus and lupus nephritis. J Rheumatol. 2010;37:45–52.

    CAS  Article  Google Scholar 

  28. 28.

    Finch DK, Ettinger R, Karnell JL, Herbst R, Sleeman MA. Effects of CXCL13 inhibition on lymphoid follicles in models of autoimmune disease. Eur J Clin Investig. 2013;43:501–9.

    CAS  Article  Google Scholar 

  29. 29.

    Herndon TM, Chen SS, Saba NS, Valdez J, Emson C, Gatmaitan M, et al. Direct in vivo evidence for increased proliferation of CLL cells in lymph nodes compared to bone marrow and peripheral blood. Leukemia. 2017;31:1340–7.

    Article  Google Scholar 

  30. 30.

    Niemann CU, Herman SE, Maric I, Gomez-Rodriguez J, Biancotto A, Chang BY, et al. Disruption of in vivo chronic lymphocytic leukemia tumor-microenvironment interactions by ibrutinib-findings from an investigator-initiated phase II study. Clin Cancer Res. 2016;22:1572–82.

    CAS  Article  Google Scholar 

  31. 31.

    Ahn IE, Underbayev C, Albitar A, Herman SE, Tian X, Maric I, et al. Clonal evolution leading to ibrutinib resistance in chronic lymphocytic leukemia. Blood. 2017;129:1469–79.

    CAS  Article  Google Scholar 

  32. 32.

    Heinig K, Gatjen M, Grau M, Stache V, Anagnostopoulos I, Gerlach K, et al. Access to follicular dendritic cells is a pivotal step in murine chronic lymphocytic leukemia B-cell activation and proliferation. Cancer Discov. 2014;4:1448–65.

    CAS  Article  Google Scholar 

  33. 33.

    Farinello D, Wozinska M, Lenti E, Genovese L, Bianchessi S, Migliori E, et al. A retinoic acid-dependent stroma-leukemia crosstalk promotes chronic lymphocytic leukemia progression. Nat Commun. 2018;9:1787.

    Article  Google Scholar 

Download references


The authors are grateful to all the patients who provided samples for this study. We thank Claire Pacelli for assistance with tissue bank. This work was supported by MD Anderson’s CLL Moonshot program (JAB), and in part by the MD Anderson Cancer Center Support Grant CA016672.

Author information




MS performed chemokine measurements, collected clinical information, analyzed the data and results, designed the figures, and wrote the paper. LX and XH performed the statistical analysis and assisted with the data interpretation. EK and AV contributed with sample collection and storage. S-SC and NC analyzed birth rate data. MJK, AF, ZE, NJ, and WGW contributed to clinical patient management. JAB designed and supervised the study, and wrote the paper together with MS. All authors reviewed the manuscript and approved the final version.

Corresponding author

Correspondence to Jan A. Burger.

Ethics declarations

Conflict of interest

JAB, NJ, and WGW received research funding from Pharmacyclics, an AbbVie company. The remaining authors declared no competing financial interests.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Sivina, M., Xiao, L., Kim, E. et al. CXCL13 plasma levels function as a biomarker for disease activity in patients with chronic lymphocytic leukemia. Leukemia (2020).

Download citation


Quick links