Tviblindi algorithm identifies branching developmental trajectories of human B‐cell development and describes abnormalities in RAG‐1 and WAS patients

European Journal of Immunology, vol. 54, pp. e2451004

Abstract

Detailed knowledge of human B-cell development is crucial for the proper interpretation of inborn errors of immunity and malignant diseases. It is of interest to understand the kinetics of protein expression changes during development, but also to properly interpret the major and possibly alternative developmental trajectories. We have investigated human samples from healthy individuals with the aim of describing all B-cell developmental trajectories. We validated a 30-parameter mass cytometry panel and demonstrated the utility of "vaevictis" visualization of B-cell developmental stages. We used the trajectory inference tool "tviblindi" to exhaustively describe all trajectories leading to all developmental ends discovered in the data. Focusing on Natural Effector B cells, we demonstrated the dynamics of expression of nuclear factors (PAX-5, TdT, Ki-67, Bcl-2), cytokine and chemokine receptors (CD127, CXCR4, CXCR5) in relation to the canonical B-cell developmental stage markers. We observed branching of the memory development, where follicular memory formation was marked by CD73 expression. Lastly, we performed an analysis of two example cases of abnormal B-cell development caused by mutations in RAG-1 and Wiskott-Aldrich syndrome gene in patients with primary immunodeficiency. In conclusion, we developed, validated, and presented a comprehensive set of tools for the investigation of B-cell development in the bone marrow compartment.

Authors 13

1. Marina Bakardjieva

   Charles University

   Affiliation as printed

   CLIP Department of Paediatric Haematology and Oncology Second Faculty of Medicine Charles University Prague Czech Republic

2. Ondřej Pelák

   Charles University

   Affiliation as printed

   CLIP Department of Paediatric Haematology and Oncology Second Faculty of Medicine Charles University Prague Czech Republic

3. Marjolein W. J. Wentink

   Erasmus MC

   Affiliation as printed

   Department of Internal Medicine Erasmus MC University Medical Center Rotterdam Rotterdam the Netherlands

4. Hana Glier

   Charles University

   Affiliation as printed

   CLIP Department of Paediatric Haematology and Oncology Second Faculty of Medicine Charles University Prague Czech Republic

5. David Novák

   Charles University · Ghent University · VIB-UGent Center for Inflammation Research

   Affiliation as printed

   CLIP Department of Paediatric Haematology and Oncology Second Faculty of Medicine Charles University Prague Czech Republic

   Data Mining and Modeling for Biomedicine Center for Inflammation Research VIB‐UGent Ghent Belgium

   Department of Applied Mathematics Computer Science and Statistics Ghent University Ghent Belgium

6. Jitka Stančíková

   Charles University

   Affiliation as printed

   CLIP Department of Paediatric Haematology and Oncology Second Faculty of Medicine Charles University Prague Czech Republic

7. Daniela Kužílková

   Charles University · University Hospital in Motol

   Affiliation as printed

   CLIP Department of Paediatric Haematology and Oncology Second Faculty of Medicine Charles University Prague Czech Republic

   Department of Paediatric Haematology and Oncology University Hospital Motol Prague Czech Republic

8. Ester Mejstříková

   Charles University · University Hospital in Motol

   Affiliation as printed

   CLIP Department of Paediatric Haematology and Oncology Second Faculty of Medicine Charles University Prague Czech Republic

   Department of Paediatric Haematology and Oncology University Hospital Motol Prague Czech Republic

9. Iga Janowska

   University of Freiburg · University Medical Center Freiburg

   Affiliation as printed

   Center for Chronic Immunodeficiency University Medical Center Freiburg Faculty of Medicine University of Freiburg Freiburg Germany

   Department of Rheumatology and Clinical Immunology Freiburg University Medical Center University of Freiburg Freiburg Germany

10. Marta Rizzi

    University of Freiburg · University Medical Center Freiburg

    Affiliation as printed

    Center for Chronic Immunodeficiency University Medical Center Freiburg Faculty of Medicine University of Freiburg Freiburg Germany

    Department of Rheumatology and Clinical Immunology Freiburg University Medical Center University of Freiburg Freiburg Germany

11. Mirjam van der Burg Aachen Department of Pediatrics Laboratory for Pediatric Immmunology Leiden University Medical Center Leiden the Netherlands

    Leiden University Medical Center

    Affiliation as printed

    Department of Pediatrics Laboratory for Pediatric Immmunology Leiden University Medical Center Leiden the Netherlands

12. Jan Stuchlý corresponding

    Charles University · University Hospital in Motol

    Affiliation as printed

    CLIP Department of Paediatric Haematology and Oncology Second Faculty of Medicine Charles University Prague Czech Republic

    Department of Paediatric Haematology and Oncology University Hospital Motol Prague Czech Republic

13. Tomáš Kalina corresponding

    Charles University · University Hospital in Motol

    Affiliation as printed

    CLIP Department of Paediatric Haematology and Oncology Second Faculty of Medicine Charles University Prague Czech Republic

    Department of Paediatric Haematology and Oncology University Hospital Motol Prague Czech Republic