Skip to main content
Advertisement

Main menu

  • Home
  • Articles
    • Newest Articles
    • Current Issue
    • Methods & Resources
    • Archive
    • Subjects
  • Collections
  • Submit
    • Submit a Manuscript
    • Author Guidelines
    • License, Copyright, Fee
    • FAQ
    • Why Submit
  • About
    • About Us
    • Editors & Staff
    • Board Members
    • Licensing and Reuse
    • Reviewer Guidelines
    • Privacy Policy
    • Advertise
    • Contact Us
    • LSA LLC
  • Alerts
  • Other Publications
    • EMBO Press
    • The EMBO Journal
    • EMBO reports
    • EMBO Molecular Medicine
    • Molecular Systems Biology
    • Rockefeller University Press
    • Journal of Cell Biology
    • Journal of Experimental Medicine
    • Journal of General Physiology
    • Cold Spring Harbor Laboratory Press
    • Genes & Development
    • Genome Research

User menu

  • My alerts

Search

  • Advanced search
Life Science Alliance
  • Other Publications
    • EMBO Press
    • The EMBO Journal
    • EMBO reports
    • EMBO Molecular Medicine
    • Molecular Systems Biology
    • Rockefeller University Press
    • Journal of Cell Biology
    • Journal of Experimental Medicine
    • Journal of General Physiology
    • Cold Spring Harbor Laboratory Press
    • Genes & Development
    • Genome Research
  • My alerts
Life Science Alliance

Advanced Search

  • Home
  • Articles
    • Newest Articles
    • Current Issue
    • Methods & Resources
    • Archive
    • Subjects
  • Collections
  • Submit
    • Submit a Manuscript
    • Author Guidelines
    • License, Copyright, Fee
    • FAQ
    • Why Submit
  • About
    • About Us
    • Editors & Staff
    • Board Members
    • Licensing and Reuse
    • Reviewer Guidelines
    • Privacy Policy
    • Advertise
    • Contact Us
    • LSA LLC
  • Alerts
  • Follow lsa Template on Twitter
Research Article
Transparent Process
Open Access

MicroRNA-155 is essential for the optimal proliferation and survival of plasmablast B cells

View ORCID ProfileGiuseppina Arbore, Tom Henley, Laura Biggins, Simon Andrews, Elena Vigorito, View ORCID ProfileMartin Turner, View ORCID ProfileRebecca Leyland  Correspondence email
Giuseppina Arbore
1Lymphocyte Signalling and Development, Babraham Institute, Cambridge, UK
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Giuseppina Arbore
Tom Henley
1Lymphocyte Signalling and Development, Babraham Institute, Cambridge, UK
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Laura Biggins
3Bioinformatics, Babraham Institute, Cambridge, UK
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Simon Andrews
3Bioinformatics, Babraham Institute, Cambridge, UK
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Elena Vigorito
1Lymphocyte Signalling and Development, Babraham Institute, Cambridge, UK
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Martin Turner
1Lymphocyte Signalling and Development, Babraham Institute, Cambridge, UK
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Martin Turner
Rebecca Leyland
1Lymphocyte Signalling and Development, Babraham Institute, Cambridge, UK
2Biomolecular Sciences Research Centre, Sheffield Hallam University, Sheffield, UK
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Rebecca Leyland
  • For correspondence: R.Leyland@shu.ac.uk
Published 16 May 2019. DOI: 10.26508/lsa.201800244
  • Article
  • Figures & Data
  • Info
  • Metrics
  • Reviewer Comments
  • PDF
Loading

Article Figures & Data

Figures

  • Supplementary Materials
  • Figure 1.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 1. miR-155 is required to sustain the plasmablast B-cell response.

    (A) A representative histogram showing HEL expression level on conjugated HEL-SRBCs (red) compared with unstained control (grey). (B) Representative flow cytometric plot showing gating strategy for SWHEL Mir155+/+ B cells at days 4.5 post immunisation, for identification of CD45.2+ donor derived HEL BCR+, B220lo plasmablast B cells or HEL BCR+, B220hi germinal centre B cells. (C) The number of SWHEL Mir155+/+ (black) or Mir155−/− (grey) HEL-specific B-cell blasts, plasmablast B cells and germinal centre B cells was calculated per 106 lymphocytes after immunisation in mice (N = 16–19 independent Mir155+/+ samples and 10–24 independent Mir155−/− samples). Data are representative of at least two independent experiments. For B-cell blast data, a Welch’s t test was used. For plasmablast and germinal centre data, P-values are from t tests using the error mean square from the ANOVA. (D) HEL-specific antibodies of the indicated immunoglobulins were measured in the serum of mice injected with SWHEL Mir155+/+ (black) or Mir155−/− (grey) B cells, at day 4.5 post immunisation with HEL-SRBCs. Red dotted line represents statistical analysis of indicated Mir155+/+ or Mir155−/− values using two-way ANOVA with Sidak’s multiple comparison test where **P < 0.01, ***P < 0.001, ****P < 0.0001.

  • Figure 2.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 2. miR-155 is essential for the optimal proliferation of plasmablast B cells.

    (A) SWHEL Mir155+/+ (black line) or Mir155−/− (grey line) HEL-binding, B220+ B cell blasts were assessed for different CFSE dilutions. (B) The percentage of CFSE+ cells in each division at days 2.5 and 3.5 post immunisation. The generation number is indicated above each data set. A two-way ANOVA with Sidak’s multiple comparison test was used. Values shown are mean ± SD. (C) Cell cycle analysis using DAPI staining was carried out on SWHEL Mir155+/+ or Mir155−/− plasmablast B cells at 3.5 and 4.5 d post immunisation. The frequency of cells in the S-G2-M phases in Mir155+/+ (black) or Mir155−/− (grey) plasmablasts is shown. A nonparametric Mann–Whitney test was used per time point. (D) Representative flow cytometry plot of DAPI staining at 4.5 d post immunisation in miR-155–sufficient or miR-155–deficient plasmablast B cells. Cells previously gated on lymphocytes and single cells. (E) EdU incorporation in SWHEL Mir155+/+ (black) or Mir155−/− (grey) plasmablast B cells at the time points indicated post immunisation. Two-way ANOVA was used. There was no significant interaction between genotype and day, but both genotype and day had significant effects P < 0.001. (F) Representative flow cytometry histograms of EdU incorporation in wild-type (WT, black line) or miR-155–deficient plasmablast B cells (black line) compared with plasmablast B cells from control mice not injected with EdU (grey) at 4 d post immunisation. Data are representative of at least two independent experiments.

  • Figure 3.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 3. miR-155 regulates survival of plasmablast B cells.

    (A) SWHEL HEL BCR+ B220lo Mir155+/+ (black) or Mir155−/− (grey) plasmablast B cells were analysed for activated caspases at the time points shown. A nonparametric Mann–Whitney test was used per time point. (B) Representative flow cytometry histograms of cleaved caspase detection in wild-type or miR-155−/− SWHEL plasmablast B cells at day 4.5 post immunisation. (C) Expression of the human Bcl2 transgene (huBCL2) in SWHEL Mir155+/+ × Bcl2 (black line) or SWHEL Mir155−/− × Bcl2 (solid grey line) plasmablast B cells, compared with negative control (single grey line). (D) Representative FACS plot showing the gating strategy for analysis of adoptively transferred SWHEL Mir155+/+ or Mir155−/− B cells expressing a human Bcl2 transgene at day 4.5 post immunisation. Plots were previously gated on lymphocytes and single cells and analysed for CD45.2 donor cells and HEL+ B220lo plasmablast B cells. HEL+ B220hi germinal centre B cells could also be visualised in all mouse strains. (E) The number of splenic SWHEL Mir155+/+ or Mir155−/− plasmablast B cells in mice with or without the expression of a human Bcl2 transgene per 106 lymphocytes. A nonparametric Kruskal–Wallis with Dunn’s multiple comparisons test was used where **P < 0.01. (F) The frequency of active caspases in miR-155–sufficient and miR-155–deficient plasmablast B cells at day 4.5 post immunisation. At least two independent experiments were carried out. Statistics calculated using nonparametric Kruksal–Wallis with Dunn’s multiple comparisons test where *P < 0.05, **P < 0.01, ****P < 0.0001. Data are representative of at least two independent experiments.

  • Figure 4.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 4. miR-155 regulates the expression of genes associated with DNA replication.

    (A) Representative gating strategy showing CD45.2+ plasmablast B cells before and after suboptimal depletion of CD45.1+ cells. After enrichment, SWHEL Mir155+/+ or Mir155−/− plasmablast B cells were sorted on a FACS Aria to more than 98% purity and analysed by microarray analysis. N = 4–5 Mir155+/+ and 8–10 Mir155−/− mice at day 4.5 post immunisation. (B) Differentially expressed (DE) genes with known functions were discovered by GOrilla analysis using the genes that were down-regulated in SWHEL Mir155−/− plasmablasts compared with SWHEL Mir155+/+ plasmablasts. (C) The mRNA abundance of down-regulated genes with reported roles in DNA replication and function in Mir155−/− plasmablasts relative to Mir155+/+ plasmablasts (dotted line) was confirmed by RT-qPCR. Expression values are normalized to HPRT, determined from three to four biological replicates from three to four independent sorting experiments using 4–5 Mir155+/+ or 8–10 Mir155−/− mice per group. (D) GOrilla analysis of the DE genes that were up-regulated in SWHEL Mir155−/− plasmablast B cells compared with SWHEL Mir155+/+ plasmablast B cells.

Supplementary Materials

  • Figures
  • Table S1 GOrilla GO enrichment analysis of down-regulated genes in miR-155–deficient plasmablasts compared with wild-type plasmablasts.

  • Table S2 GOrilla GO enrichment analysis of up-regulated genes in miR-155–deficient plasmablasts compared with wild-type plasmablasts.

  • Table S3 Primer sequences used for RT-qPCR.

PreviousNext
Back to top
Download PDF
Article Alerts
Sign In to Email Alerts with your Email Address
Email Article

Thank you for your interest in spreading the word on Life Science Alliance.

NOTE: We only request your email address so that the person you are recommending the page to knows that you wanted them to see it, and that it is not junk mail. We do not capture any email address.

Enter multiple addresses on separate lines or separate them with commas.
MicroRNA-155 is essential for the optimal proliferation and survival of plasmablast B cells
(Your Name) has sent you a message from Life Science Alliance
(Your Name) thought you would like to see the Life Science Alliance web site.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Citation Tools
MicroRNA-155 is essential for the plasmablast response
Giuseppina Arbore, Tom Henley, Laura Biggins, Simon Andrews, Elena Vigorito, Martin Turner, Rebecca Leyland
Life Science Alliance May 2019, 2 (3) e201800244; DOI: 10.26508/lsa.201800244

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Share
MicroRNA-155 is essential for the plasmablast response
Giuseppina Arbore, Tom Henley, Laura Biggins, Simon Andrews, Elena Vigorito, Martin Turner, Rebecca Leyland
Life Science Alliance May 2019, 2 (3) e201800244; DOI: 10.26508/lsa.201800244
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Tweet Widget
  • Facebook Like
Issue Cover

In this Issue

Volume 2, No. 3
June 2019
  • Table of Contents
  • Cover (PDF)
  • About the Cover
  • Masthead (PDF)
Advertisement

Jump to section

  • Article
    • Abstract
    • Introduction
    • Results and Discussion
    • Materials and Methods
    • Acknowledgements
    • References
  • Figures & Data
  • Info
  • Metrics
  • Reviewer Comments
  • PDF

Subjects

  • Immunology

Related Articles

  • No related articles found.

Cited By...

  • No citing articles found.
  • Google Scholar

More in this TOC Section

  • NLRP3 controls ATM activation
  • RIF1 stabilizes replication domains
  • Chromatin context-dependent interferon response
Show more Research Article

Similar Articles

EMBO Press LogoRockefeller University Press LogoCold Spring Harbor Logo

Content

  • Home
  • Newest Articles
  • Current Issue
  • Archive
  • Subject Collections

For Authors

  • Submit a Manuscript
  • Author Guidelines
  • License, copyright, Fee

Other Services

  • Alerts
  • Twitter
  • RSS Feeds

More Information

  • Editors & Staff
  • Reviewer Guidelines
  • Feedback
  • Licensing and Reuse
  • Privacy Policy

ISSN: 2575-1077
© 2023 Life Science Alliance LLC

Life Science Alliance is registered as a trademark in the U.S. Patent and Trade Mark Office and in the European Union Intellectual Property Office.