Abstract

Differentiation of B cells into antibody-secreting cells (ASCs) is a key process to generate protective humoral immunity. A detailed understanding of the cues controlling ASC differentiation is important to devise strategies to modulate antibody formation. Here, we dissected differentiation trajectories of human naive B cells into ASCs using single-cell RNA sequencing. By comparing transcriptomes of B cells at different stages of differentiation from an in vitro model with ex vivo B cells and ASCs, we uncovered a novel pre-ASC population present ex vivo in lymphoid tissues. For the first time, a germinal-center-like population is identified in vitro from human naive B cells and possibly progresses into a memory B cell population through an alternative route of differentiation, thus recapitulating in vivo human GC reactions. Our work allows further detailed characterization of human B cell differentiation into ASCs or memory B cells in both healthy and diseased conditions.

Data availability

All raw fastq files and digital gene expression matrixes (DGE) are available at the Gene Expression Omnibus (GEO) repository under accession no. GSE214265.

The following data sets were generated
The following previously published data sets were used

Article and author information

Author details

  1. Niels JM Verstegen

    Department of Immunopathology, Sanquin, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5732-4979
  2. Sabrina Pollastro

    Department of Immunopathology, Sanquin, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0950-7583
  3. Peter-Paul A Unger

    Department of Immunopathology, Sanquin, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  4. Casper Marsman

    Department of Immunopathology, Sanquin, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  5. George Elias

    Department of Immunopathology, Sanquin, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8419-9544
  6. Tineke Jorritsma

    Department of Immunopathology, Sanquin, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  7. Marij Streutker

    Department of Immunopathology, Sanquin, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  8. kevin bassler

    Genomics and Immunoregulation, University of Bonn, Bonn, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4780-372X
  9. Kristian Haendler

    Genomics and Immunoregulation, University of Bonn, Bonn, Germany
    Competing interests
    The authors declare that no competing interests exist.
  10. Theo Rispens

    Department of Immunopathology, Sanquin, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  11. Joachim L Schultze

    Genomics and Immunoregulation, University of Bonn, Bonn, Germany
    Competing interests
    The authors declare that no competing interests exist.
  12. Anja ten Brinke

    Department of Immunopathology, Sanquin, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  13. Marc Beyer

    Genomics and Immunoregulation, University of Bonn, Bonn, Germany
    Competing interests
    The authors declare that no competing interests exist.
  14. Marieke SM van Ham

    Department of Immunopathology, Sanquin, Amsterdam, Netherlands
    For correspondence
    m.vanham@sanquin.nl
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1999-9494

Funding

LSBR (1609)

  • Marieke SM van Ham

PPOC (17-34/ L2263)

  • Marieke SM van Ham

German research foundation (272482170)

  • Marc Beyer

German research foundation (SFB1454-432325352)

  • Marc Beyer

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Murim Choi, Seoul National University, Republic of Korea

Ethics

Human subjects: All the healthy donors provided written informed consent following the protocol of the local institutional review board, the Medical Ethics Committee of Sanquin Blood Supply, and conforms to the principles of the Declaration of Helsinki.

Version history

  1. Received: September 20, 2022
  2. Preprint posted: October 5, 2022 (view preprint)
  3. Accepted: February 24, 2023
  4. Accepted Manuscript published: March 2, 2023 (version 1)
  5. Version of Record published: March 10, 2023 (version 2)

Copyright

© 2023, Verstegen et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 3,981
    views
  • 579
    downloads
  • 4
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Niels JM Verstegen
  2. Sabrina Pollastro
  3. Peter-Paul A Unger
  4. Casper Marsman
  5. George Elias
  6. Tineke Jorritsma
  7. Marij Streutker
  8. kevin bassler
  9. Kristian Haendler
  10. Theo Rispens
  11. Joachim L Schultze
  12. Anja ten Brinke
  13. Marc Beyer
  14. Marieke SM van Ham
(2023)
Single-cell analysis reveals dynamics of human B cell differentiation and identifies novel B and antibody-secreting cell intermediates
eLife 12:e83578.
https://doi.org/10.7554/eLife.83578

Share this article

https://doi.org/10.7554/eLife.83578

Further reading

    1. Cell Biology
    2. Chromosomes and Gene Expression
    Lucie Crhak Khaitova, Pavlina Mikulkova ... Karel Riha
    Research Article

    Heat stress is a major threat to global crop production, and understanding its impact on plant fertility is crucial for developing climate-resilient crops. Despite the known negative effects of heat stress on plant reproduction, the underlying molecular mechanisms remain poorly understood. Here, we investigated the impact of elevated temperature on centromere structure and chromosome segregation during meiosis in Arabidopsis thaliana. Consistent with previous studies, heat stress leads to a decline in fertility and micronuclei formation in pollen mother cells. Our results reveal that elevated temperature causes a decrease in the amount of centromeric histone and the kinetochore protein BMF1 at meiotic centromeres with increasing temperature. Furthermore, we show that heat stress increases the duration of meiotic divisions and prolongs the activity of the spindle assembly checkpoint during meiosis I, indicating an impaired efficiency of the kinetochore attachments to spindle microtubules. Our analysis of mutants with reduced levels of centromeric histone suggests that weakened centromeres sensitize plants to elevated temperature, resulting in meiotic defects and reduced fertility even at moderate temperatures. These results indicate that the structure and functionality of meiotic centromeres in Arabidopsis are highly sensitive to heat stress, and suggest that centromeres and kinetochores may represent a critical bottleneck in plant adaptation to increasing temperatures.

    1. Cell Biology
    Wan-ping Yang, Mei-qi Li ... Qian-qian Luo
    Research Article

    High-altitude polycythemia (HAPC) affects individuals living at high altitudes, characterized by increased red blood cells (RBCs) production in response to hypoxic conditions. The exact mechanisms behind HAPC are not fully understood. We utilized a mouse model exposed to hypobaric hypoxia (HH), replicating the environmental conditions experienced at 6000 m above sea level, coupled with in vitro analysis of primary splenic macrophages under 1% O2 to investigate these mechanisms. Our findings indicate that HH significantly boosts erythropoiesis, leading to erythrocytosis and splenic changes, including initial contraction to splenomegaly over 14 days. A notable decrease in red pulp macrophages (RPMs) in the spleen, essential for RBCs processing, was observed, correlating with increased iron release and signs of ferroptosis. Prolonged exposure to hypoxia further exacerbated these effects, mirrored in human peripheral blood mononuclear cells. Single-cell sequencing showed a marked reduction in macrophage populations, affecting the spleen’s ability to clear RBCs and contributing to splenomegaly. Our findings suggest splenic ferroptosis contributes to decreased RPMs, affecting erythrophagocytosis and potentially fostering continuous RBCs production in HAPC. These insights could guide the development of targeted therapies for HAPC, emphasizing the importance of splenic macrophages in disease pathology.