Characterisation of the immune repertoire of a humanised transgenic mouse through immunophenotyping and high-throughput sequencing
- University of Oxford, United Kingdom
- Kymab, United Kingdom
- University of Zurich, Switzerland
- 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).
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Paired VH:VL IGM sequences from 22 naïve KymiceZenodo, 10.5281/zenodo.7474232.
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Pre-processed IgH repertoire sequencing data from BioProject PRJNA748239Zenodo, doi:10.5281/zenodo.5155565.
Article and author information
Author details
Valentin von Niederhäusern
Aleksandr Kovaltsuk
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.
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).
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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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
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