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Home › Centrum für Chronische Immundefizienz (CCI) › deutsch › Centrum für Chronische Immundefizienz (CCI) › Forschung/Studien › Forschungsgruppen › Bärbel Keller

Alternative B cell differentiation and signaling in immune dysregulation

Head: Dr. Bärbel Keller

Off track - The alternative differentiation of T-bethighCD21low B cells

Summary

The humoral immune response plays a crucial part in the defense of the adaptive immune system against especially extracellular pathogens, but also viral infections as recently demonstrated by COVID19. In addition, B-cell mediated responses play also detrimental roles in certain autoimmune disorders like systemic lupus erythematosus (SLE), ANCA-associated vasculitis, rheumatoid arthritis and others. We have discovered an alternative differentiation pathway of activated B cells leading to the expansion of T-bethighCD21low B cells.

We have described these cells first in SLE and common variable immunodeficiency (CVID) (1-3) while others have found these cells in viremic HIV patients. Later we and others could demonstrate the expansion of this population also in other autoimmune disorders and viral as well as parasitic infections (4). While a temporary expansion can be observed after vaccination against influenza and SARS-CoV-2, a persistent expansion is seen in some patients with chronic active, TH1-dominated immune stimulation (5), especially at sites of inflammation like the joint in patients with rheumatoid arthritis or the bronchoalveolar lavage of CVID patients with interstitial lung disease (6, 7). While this phenotype can present during different stages of B-cell development it is strongly overlapping indicative of an activated stage with the characteristic expression of markers like several inhibitory receptors, FAS, CD86 and CD11c (4). A severely altered signaling network (4, 8-11) is associated with a distinct gene expression profile and distribution of open chromatin regions of these cells and the transcription factor T-bet has been identified as a hallmark of this population (12). Extensive phenotyping of monogenetic disorders has identified T-cell derived signals like CD40L-CD40, IL21, IFNg crucial determinants for the differentiation of this unique B cell phenotype (12). Future research will address the transcriptional network determining their fate in more detail, their role in virus infection as well as in autoimmunity.

THE CELL
THE SIGNALING
THE ENHANCER REPERTOIRE AND TRANSCRIPTOME

Methods

  • Deep phenotyping of the immune system
  • Cell culture studies
  • Signaling
  • RNAseq
  • ATACseq

Cooperation Partner

  • Stuart Tangye (Sydney, Australia)
  • Melanie Börries (IMBI, Freiburg)

Funding

  • DFG TRR130 (2013-2022)

Publications

  1. Wehr, C., H. Eibel, M. Masilamani, H. Illges, M. Schlesier, H. H. Peter, and K. Warnatz. 2004. A new CD21low B cell population in the peripheral blood of patients with SLE. Clinical Immunology 113: 161-171.
  2. Warnatz, K., C. Wehr, R. Drager, S. Schmidt, H. Eibel, M. Schlesier, and H. H. Peter. 2002. Expansion of CD19(hi)CD21(lo/neg) B cells in common variable immunodeficiency (CVID) patients with autoimmune cytopenia. Immunobiology 206: 502-513.
  3. Warnatz, K., A. Denz, R. Drager, M. Braun, C. Groth, G. Wolff-Vorbeck, H. Eibel, M. Schlesier, and H. H. Peter. 2002. Severe deficiency of switched memory B cells (CD27(+)IgM(-)IgD(-)) in subgroups of patients with common variable immunodeficiency: a new approach to classify a heterogeneous disease. Blood 99: 1544-1551.
  4. Freudenhammer, M., R. E. Voll, S. C. Binder, B. Keller, and K. Warnatz. 2020. Naive- and Memory-like CD21(low) B Cell Subsets Share Core Phenotypic and Signaling Characteristics in Systemic Autoimmune Disorders. J Immunol 205: 2016-2025.
  5. Unger, S., M. Seidl, P. van Schouwenburg, M. Rakhmanov, A. Bulashevska, N. Frede, B. Grimbacher, J. Pfeiffer, K. Schrenk, L. Munoz, L. Hanitsch, I. Stumpf, F. Kaiser, O. Hausmann, F. Kollert, S. Goldacker, M. van der Burg, B. Keller, and K. Warnatz. 2018. TH1 phenotype of T follicular helper cells indicates an IFNgamma-associated immune dysregulation in CD21low CVID patients. J Allergy Clin Immunol 141: 730-740.
  6. Rakhmanov, M., B. Keller, S. Gutenberger, C. Foerster, M. Hoenig, G. Driessen, B. M. van der, J. J. van Dongen, E. Wiech, M. Visentini, I. Quinti, A. Prasse, N. Voelxen, U. Salzer, S. Goldacker, P. Fisch, H. Eibel, K. Schwarz, H. H. Peter, and K. Warnatz. 2009. Circulating CD21low B cells in common variable immunodeficiency resemble tissue homing, innate-like B cells. Proc.Natl.Acad.Sci.U.S.A. 106(32): 13451-13456.
  7. Friedmann, D., S. Unger, B. Keller, M. Rakhmanov, S. Goldacker, G. Zissel, B. C. Frye, J. C. Schupp, A. Prasse, and K. Warnatz. 2020. Bronchoalveolar Lavage Fluid Reflects a T(H)1-CD21(low) B-Cell Interaction in CVID-Related Interstitial Lung Disease. Frontiers in immunology 11: 616832.
  8. Harder, I., M. Münchhalfen, G. Andrieux, M. Boerries, B. Grimbacher, H. Eibel, M. E. Maccari, S. Ehl, J. Wienands, J. Jellusova, K. Warnatz, and B. Keller. 2022. Dysregulated PI3K Signaling in B Cells of CVID Patients. Cells 11: 464.
  9. Keller, B., Z. Cseresnyes, I. Stumpf, C. Wehr, M. Fliegauf, A. Bulashevska, S. Usadel, B. Grimbacher, M. Rizzi, H. Eibel, R. Niesner, and K. Warnatz. 2017. Disturbed canonical nuclear factor of kappa light chain signaling in B cells of patients with common variable immunodeficiency. J Allergy Clin Immunol 139: 220-231.e228.
  10. Keller, B., I. Stumpf, V. Strohmeier, S. Usadel, E. Verhoeyen, H. Eibel, and K. Warnatz. 2017. High SYK Expression Drives Constitutive Activation of CD21low B Cells. J Immunol 198: 4285-4292.
  11. Foerster, C., N. Voelxen, M. Rakhmanov, B. Keller, S. Gutenberger, S. Goldacker, J. Thiel, S. Feske, H. H. Peter, and K. Warnatz. 2010. B cell receptor-mediated calcium signaling is impaired in B lymphocytes of type Ia patients with common variable immunodeficiency. Journal of Immunology 184(12): 7305-7313.
  12. Keller, B., V. Strohmeier, I. Harder, S. Unger, K. J. Payne, G. Andrieux, M. Boerries, P. T. Felixberger, J. J. M. Landry, A. Nieters, A. Rensing-Ehl, U. Salzer, N. Frede, S. Usadel, R. Elling, C. Speckmann, I. Hainmann, E. Ralph, K. Gilmour, M. W. J. Wentink, M. van der Burg, H. S. Kuehn, S. D. Rosenzweig, U. Kolsch, H. von Bernuth, P. Kaiser-Labusch, F. Gothe, S. Hambleton, A. D. Vlagea, A. Garcia Garcia, L. Alsina, G. Markelj, T. Avcin, J. Vasconcelos, M. Guedes, J. Y. Ding, C. L. Ku, B. Shadur, D. T. Avery, N. Venhoff, J. Thiel, H. Becker, L. Erazo-Borras, C. M. Trujillo-Vargas, J. L. Franco, C. Fieschi, S. Okada, P. E. Gray, G. Uzel, J. L. Casanova, M. Fliegauf, B. Grimbacher, H. Eibel, S. Ehl, R. E. Voll, M. Rizzi, P. Stepensky, V. Benes, C. S. Ma, C. Bossen, S. G. Tangye, and K. Warnatz. 2021. The expansion of human T-bet(high)CD21(low) B cells is T cell dependent. Science immunology 6: eabh0891.

 

 

 

 

Blocked roads – Altered signaling in B and T cells – monogenic defects or just dysregulation, with a focus on NF-κB

Summary

Signaling downstream of the immune receptor is crucial for the cellular processes and integrity of the adaptive immune system. In collaboration with Polina Stepensky (Hadassah, Jerusalem) and colleagues from Palestine, we have described the first patient with CARD11 deficiency presenting with profound combined immunodeficiency, with abrogated canonical NF-κB signaling downstream of the T and B cell receptor and a severe block in B cell differentiation (1). Additionally, we investigated patients with combined immunodeficiency due to inborn errors of immunity (IEI) affecting LAT, with unexpectedly preserved canonical NF-κB as well as Ca2+ signaling (2); in LCK with broadly affecting TCR signaling but a potentially preserved PI3K pathway (3); and observed severely disturbed antigen receptor-mediated canonical NF-κB signaling in BCL10 deficiency (4). Moreover we uncovered reduced canonical NF-κB signaling in B cells from patients with CIN85 deficiency, resulting in hypogammaglobulinemia (5). Aberrant antigen receptor signaling affects lymphocyte development, activation and differentiation and slightly altered signaling hubs in these patients affect the T and B cell compartment (6). Moreover, signaling transduction and intensity varies in the different T and B cell subpopulations. Our goal is to elucidate these influence of specific signaling defect on human B and CD4 T cell development and differentiation.

Beside our analysis in patients with IEI, we recently have identified disturbed BCR signaling in patients with common variable immunodeficiency (CVID) without defined monogenic defects. Despite normal BCR proximal signaling in the CD21pos naïve B cell compartment (7), Ca2+ signaling (8) as well as canonical NF-κB signaling (9) was reduced in patients with TH1-linked immune dysregulation and an accumulation of T-bethighCD21low B cells but less so in patients with mere susceptibility to infections. Similarly, PI3K signaling was reduced in B cells from these complicated CVID patients (10). We hypothesize, that factors causing or originating from the prevailing immune dysregulation compromise the integrity of key signaling pathways downstream of BCR and TCR signaling.

CARD11 DEFICIENCY
MONOGENIC DEFECTS IN ANTIGEN RECEPTOR SIGNALING
BCR SIGNALING DEFECTS SECONDARY TO IMMUNE DYSREGULATION

Methods

  • Deep phenotyping of the immune system
  • Signaling assays
  • Cell culture studies
  • RNAseq
  • Molecular cloning
  • Lentiviral Transduction

Cooperation Partner

  • Polina Stepensky (Jerusalem Israel) Sydney, Australia)
  • Melanie Börries (IMBI, Freiburg)
  • Jürgen Wienands (Göttingen)

Publications

1. Stepensky P, Keller B, Buchta M, Kienzler AK, Elpeleg O, Somech R, et al. Deficiency of caspase recruitment domain family, member 11 (CARD11), causes profound combined immunodeficiency in human subjects. J Allergy Clin Immunol. 2013;131(2):477-85 e1.

2. Keller B, Zaidman I, Yousefi OS, Hershkovitz D, Stein J, Unger S, et al. Early onset combined immunodeficiency and autoimmunity in patients with loss-of-function mutation in LAT. J Exp Med. 2016;213(7):1185-99.

3. Keller B, Kfir-Erenfeld S, Matusewicz P, Hartl F, Lev A, Lee YN, et al. Combined Immunodeficiency Caused by a Novel Nonsense Mutation in LCK. Journal of clinical immunology. 2023;44(1):4.

4. Salou S, Voelkl S, Keller B, Ehl S, Naumann-Bartsch N. BCL10 Deficiency Presenting as Severe Combined Immunodeficiency Escaping Newborn Screening. Journal of clinical immunology. 2023;44(1):37.

5. Keller B, Shoukier M, Schulz K, Bhatt A, Heine I, Strohmeier V, et al. Germline deletion of CIN85 in humans with X chromosome-linked antibody deficiency. J Exp Med. 2018;215(5):1327-36.

6. Keller B, Strohmeier V, Harder I, Unger S, Payne KJ, Andrieux G, et al. The expansion of human T-bet(high)CD21(low) B cells is T cell dependent. Science immunology. 2021;6(64):eabh0891.

7. Keller B, Stumpf I, Strohmeier V, Usadel S, Verhoeyen E, Eibel H, et al. High SYK Expression Drives Constitutive Activation of CD21(low) B Cells. J Immunol. 2017;198(11):4285-92.

8. Foerster C, Voelxen N, Rakhmanov M, Keller B, Gutenberger S, Goldacker S, et al. B cell receptor-mediated calcium signaling is impaired in B lymphocytes of type Ia patients with common variable immunodeficiency. J Immunol. 2010;184(12):7305-13.

9. Keller B, Cseresnyes Z, Stumpf I, Wehr C, Fliegauf M, Bulashevska A, et al. Disturbed canonical nuclear factor of kappa light chain signaling in B cells of patients with common variable immunodeficiency. J Allergy Clin Immunol. 2016.

10. Harder I, Munchhalfen M, Andrieux G, Boerries M, Grimbacher B, Eibel H, et al. Dysregulated PI3K Signaling in B Cells of CVID Patients. Cells. 2022;11(3).

 

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