An Essential Role of UBXN3B in B Lymphopoiesis

  1. Tingting Geng
  2. Duomeng Yang
  3. Tao Lin
  4. Andrew G. Harrison
  5. Binsheng Wang
  6. Blake Torrance
  7. Kepeng Wang
  8. Yanlin Wang
  9. Long Yang
  10. Laura Haynes
  11. Gong Cheng
  12. Anthony T. Vella
  13. Erol Fikrig
  14. Penghua Wang

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Abstract

Hematopoiesis is finely regulated to enable timely production of the right numbers and types of mature immune cells to maintain tissue homeostasis. Dysregulated hematopoiesis may compromise antiviral immunity and/or exacerbate immunopathogenesis. Herein, we report an essential role of UBXN3B in maintenance of hematopoietic homeostasis and restriction of immunopathogenesis during respiratory viral infection. Ubxn3b deficient ( Ubxn3b −/− ) mice are highly vulnerable to SARS-CoV-2 and influenza A infection, characterized by more severe lung immunopathology, lower virus-specific IgG, significantly fewer B cells, but more myeloid cells than Ubxn3b +/+ littermates. This aberrant immune compartmentalization is recapitulated in uninfected Ubxn3b −/− mice. Mechanistically, UBXN3B controls precursor B-I (pre-BI) transition to pre-BII and subsequent proliferation in a cell-intrinsic manner, by maintaining BLNK protein stability and pre-BCR signaling. These results reveal an essential role of UBXN3B for the early stage of B cell development.

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  1. ScreenIT

    SciScore for 10.1101/2021.03.04.433919: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    Institutional Review Board StatementIACUC: The animal protocols were approved by the Institutional Animal Care and Use Committee at the University of Connecticut and Yale University.
    RandomizationAll animal results were included and no method of randomization was applied.
    Blindingnot detected.
    Power AnalysisData Acquisition and Statistical Analysis: The sample size chosen for our animal experiments in this study was estimated according to our prior experience in similar sets of experiments and power analysis calculations (http://isogenic.info/html/power_analysis.html).
    Sex as a biological variablenot detected.
    Cell Line AuthenticationContamination: We routinely added MycoZAP (Lonza Group, Basel, Switzerland) to cell cultures prevent mycoplasma contamination.

    Table 2: Resources

    Antibodies
    SentencesResources
    Antibodies, Cell lines and Viruses: Human embryonic kidney 293 cells transformed with T antigen of SV40 (HEK293T, # CRL-3216) and Vero cells (monkey kidney epithelial cells, # CCL-81) were purchased from American Type Culture Collection (ATCC) (Manassas, VA20110, USA).
    CCL-81
    suggested: None
    Cells were suspended in FACS buffer and stained for 30 min at 4 °C with the desired antibody cocktails (Biolegend, San Diego, CA, US) of APC-Fire 750-anti CD11b (Cat. # 101261, clone M1/70), Alexa Fluor 700-anti Ly-6G (Cat. # 127621, clone 1A8)
    CD11b
    suggested: (BioLegend Cat# 101261, RRID:AB_2572121)
    Ly-6G
    suggested: (BioLegend Cat# 127621, RRID:AB_10640452)
    Experimental Models: Cell Lines
    SentencesResources
    Antibodies, Cell lines and Viruses: Human embryonic kidney 293 cells transformed with T antigen of SV40 (HEK293T, # CRL-3216) and Vero cells (monkey kidney epithelial cells, # CCL-81) were purchased from American Type Culture Collection (ATCC) (Manassas, VA20110, USA).
    HEK293T
    suggested: ATCC Cat# CRL-3216, RRID:CVCL_0063)
    Concentration of SARS-CoV-2: The virus was grown in Vero cells for 72hrs, and the culture medium was cleared by brief centrifugation.
    Vero
    suggested: None
    Experimental Models: Organisms/Strains
    SentencesResources
    1-6 Supplemental Movie 1: SARS-CoV-2-infected Ubxn3b+/+ mice on day 2 after infection.
    Ubxn3b+/+
    suggested: None
    Supplemental Movie 2: SARS-CoV-2-infected Ubxn3b-/- mice on day 2 after infection.
    Ubxn3b-/-
    suggested: None
    Software and Algorithms
    SentencesResources
    Data were analyzed using the FlowJo software.
    FlowJo
    suggested: (FlowJo, RRID:SCR_008520)
    All data were analyzed with a GraphPad Prism software by non-parametric Mann-Whitney test or two-tailed Student’s t-test depending on the data distribution.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)

    Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

    Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

    Results from TrialIdentifier: No clinical trial numbers were referenced.

    Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

    Results from JetFighter: Please consider improving the rainbow (“jet”) colormap(s) used on page 33. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.

    Results from rtransparent:
    • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
    • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
    • No protocol registration statement was detected.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

  2. Version published to 10.1101/2021.03.04.433919 on bioRxiv