Characterisation of the immune repertoire of a humanised transgenic mouse through immunophenotyping and high-throughput sequencing

  1. Eve Richardson
  2. Špela Binter
  3. Miha Kosmac
  4. Marie Ghraichy
  5. Valentin von Niederhäusern
  6. Aleksandr Kovaltsuk
  7. Jacob D Galson
  8. Johannes Trück
  9. Dominic F Kelly
  10. Charlotte M Deane  Is a corresponding author
  11. Paul Kellam
  12. Simon J Watson  Is a corresponding author
  1. University of Oxford, United Kingdom
  2. Kymab, United Kingdom
  3. University of Zurich, Switzerland
  4. Alchemab Therapeutics Ltd, United Kingdom

Abstract

Immunoglobulin loci-transgenic animals are widely used in antibody discovery and increasingly in vaccine response modelling. In this study, we phenotypically characterised B-cell populations from the Intelliselect® Transgenic mouse (Kymouse) demonstrating full B-cell development competence. Comparison of the naïve B-cell receptor (BCR) repertoires of Kymice BCRs, naïve human, and murine BCR repertoires revealed key differences in germline gene usage and junctional diversification. These differences result in Kymice having CDRH3 length and diversity intermediate between mice and humans. To compare the structural space explored by CDRH3s in each species’ repertoire, we used computational structure prediction to show that Kymouse naïve BCR repertoires are more human-like than mouse-like in their predicted distribution of CDRH3 shape. Our combined sequence and structural analysis indicates that the naïve Kymouse BCR repertoire is diverse with key similarities to human repertoires, while immunophenotyping confirms that selected naïve B-cells are able to go through complete development.

Data availability

The processed paired sequencing data is available at doing 10.5281/zenodo.7474232 and the processed bulk VH sequence data has been deposited in the Observed Antibody Space (http://opig.stats.ox.ac.uk/webapps/oas/oas). Immunophenotyping data is available in the FlowRepository (https://flowrepository.org/) under IDs FR-FCM-Z5LB (bone marrow samples) and FR-FCM-Z5LC (splenocyte and lymph node samples).

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

Article and author information

Author details

  1. Eve Richardson

    Department of Statistics, University of Oxford, Oxford, United Kingdom
    Competing interests
    Eve Richardson, Eve Richardson receives funding from Kymab Ltd..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5499-6283
  2. Špela Binter

    Kymab, Cambridge, United Kingdom
    Competing interests
    Špela Binter, Špela Binter is an employee of Kymab Ltd..
  3. Miha Kosmac

    Kymab, Cambridge, United Kingdom
    Competing interests
    Miha Kosmac, Miha Kosmac was an employee of Kymab within the last three years..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5372-7233
  4. Marie Ghraichy

    Division of Immunology, University of Zurich, Zurich, Switzerland
    Competing interests
    No competing interests declared.
  5. Valentin von Niederhäusern

    Division of Immunology, University of Zurich, Zurich, Switzerland
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9150-2361
  6. Aleksandr Kovaltsuk

    Department of Statistics, University of Oxford, Oxford, United Kingdom
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3806-8302
  7. Jacob D Galson

    Alchemab Therapeutics Ltd, London, United Kingdom
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4916-800X
  8. Johannes Trück

    Division of Immunology, University of Zurich, Zurich, Switzerland
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0418-7381
  9. Dominic F Kelly

    Department of Paediatrics, University of Oxford, Oxford, United Kingdom
    Competing interests
    No competing interests declared.
  10. Charlotte M Deane

    Department of Statistics, University of Oxford, Oxford, United Kingdom
    For correspondence
    deane@stats.ox.ac.uk
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1388-2252
  11. Paul Kellam

    Kymab, Cambridge, United Kingdom
    Competing interests
    Paul Kellam, Paul Kellam is an employee of Kymab Ltd..
  12. Simon J Watson

    Kymab, Cambridge, United Kingdom
    For correspondence
    Simon.Watson@rqbiotechnology.com
    Competing interests
    Simon J Watson, Simon Watson is an employee of Kymab Ltd..

Funding

Bill & Melinda Gates Foundation (OPP1159947)

  • Paul Kellam

Medical Research Council (MR/R015708/1)

  • Eve Richardson

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

Reviewing Editor

  1. Ranjan Sen, National Institute on Aging, United States

Ethics

Animal experimentation: This study was carried out under Project Licenses 70/8718 issued by the UK Government Home Office under Animal (Scientific Procedures) Act (A(SP)A), 1986, incorporating Directive 2010/63/EU of the European Parliament, and with the approval of the Sanger Institute Animal Welfare and Ethical Review Body. The Institute complied with the Code of Practice issued by the UK Government which aids compliance with the A(SP)A. The Institute has a PHS assurance F16-00128 (WTSI).

Version history

  1. Preprint posted: June 29, 2022 (view preprint)
  2. Received: July 5, 2022
  3. Accepted: March 26, 2023
  4. Accepted Manuscript published: March 27, 2023 (version 1)
  5. Version of Record published: April 19, 2023 (version 2)

Copyright

© 2023, Richardson 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.

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  1. Eve Richardson
  2. Špela Binter
  3. Miha Kosmac
  4. Marie Ghraichy
  5. Valentin von Niederhäusern
  6. Aleksandr Kovaltsuk
  7. Jacob D Galson
  8. Johannes Trück
  9. Dominic F Kelly
  10. Charlotte M Deane
  11. Paul Kellam
  12. Simon J Watson
(2023)
Characterisation of the immune repertoire of a humanised transgenic mouse through immunophenotyping and high-throughput sequencing
eLife 12:e81629.
https://doi.org/10.7554/eLife.81629

Share this article

https://doi.org/10.7554/eLife.81629

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