TCR transgenic clone selection guided by immune receptor analysis and single cell RNA expression of polyclonal responders
- Author
- Nincy Debeuf (UGent) , Sahine Lameire (UGent) , Manon Vanheerswynghels (UGent) , Julie Deckers (UGent) , Caroline De Wolf (UGent) , Wendy Toussaint (UGent) , Rein Verbeke (UGent) , Kevin Verstaen (UGent) , Hamida Hammad (UGent) , Stijn Vanhee (UGent) and Bart Lambrecht (UGent)
- Organization
- Project
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- GM-CSF as crucial rheostat of the allergic airways
- Understanding and modulating the pro-inflammatory activity of lipid nanoparticles (LNP) in the COVID-19 mRNA vaccines: towards the development of more effective LNP formulations.
- Uncovering a role for B cells in allergy risk and protection in early life.
- Understanding the division of labour between local and systemic humoral responses in allergic airway disease.
- An integrated approach to unravel eosinophil function in chronic eosinophilic disorders
- Understanding the immunological basis of persistent airway obstruction in asthma
- Abstract
- Since the precursor frequency of naïve T cells is extremely low, investigating the early steps of antigen-specific T cell activation is challenging. To overcome this detection problem, adoptive transfer of a cohort of T cells purified from T cell receptor (TCR) transgenic donors has been extensively used but is not readily available for emerging pathogens. Constructing TCR transgenic mice from T cell hybridomas is a labor-intensive and sometimes erratic process, since the best clones are selected based on antigen-induced CD69 upregulation or IL-2 production in vitro, and TCR chains are PCR-cloned into expression vectors. Here, we exploited the rapid advances in single cell sequencing and TCR repertoire analysis to select the best clones without hybridoma selection, and generated CORSET8 mice (CORona Spike Epitope specific CD8 T cell), carrying a TCR specific for the Spike protein of SARS-CoV-2. Implementing newly created DALI software for TCR repertoire analysis in single cell analysis enabled the rapid selection of the ideal responder CD8 T cell clone, based on antigen reactivity, proliferation and immunophenotype in vivo. In contrast, a traditional method based on hybridoma technology was unsuccessful. Identified TCR sequences were inserted as synthetic DNA into an expression vector and transgenic CORSET8 donor mice were created. After immunization with Spike/CpG-motifs, mRNA vaccination or SARS-CoV2 infection, CORSET8 T cells strongly proliferated and showed signs of T cell activation. Thus, a combination of TCR repertoire analysis and scRNA immunophenotyping allowed rapid selection of antigen-specific TCR sequences that can be used to generate TCR transgenic mice.
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Citation
Please use this url to cite or link to this publication: http://hdl.handle.net/1854/LU-01J19P07290S42DTWG26EZTZDB
- MLA
- Debeuf, Nincy, et al. “TCR Transgenic Clone Selection Guided by Immune Receptor Analysis and Single Cell RNA Expression of Polyclonal Responders.” BIORXIV, 2024, doi:10.7554/elife.98344.1.
- APA
- Debeuf, N., Lameire, S., Vanheerswynghels, M., Deckers, J., De Wolf, C., Toussaint, W., … Lambrecht, B. (2024). TCR transgenic clone selection guided by immune receptor analysis and single cell RNA expression of polyclonal responders. https://doi.org/10.7554/elife.98344.1
- Chicago author-date
- Debeuf, Nincy, Sahine Lameire, Manon Vanheerswynghels, Julie Deckers, Caroline De Wolf, Wendy Toussaint, Rein Verbeke, et al. 2024. “TCR Transgenic Clone Selection Guided by Immune Receptor Analysis and Single Cell RNA Expression of Polyclonal Responders.” BIORXIV. https://doi.org/10.7554/elife.98344.1.
- Chicago author-date (all authors)
- Debeuf, Nincy, Sahine Lameire, Manon Vanheerswynghels, Julie Deckers, Caroline De Wolf, Wendy Toussaint, Rein Verbeke, Kevin Verstaen, Hamida Hammad, Stijn Vanhee, and Bart Lambrecht. 2024. “TCR Transgenic Clone Selection Guided by Immune Receptor Analysis and Single Cell RNA Expression of Polyclonal Responders.” BIORXIV. doi:10.7554/elife.98344.1.
- Vancouver
- 1.Debeuf N, Lameire S, Vanheerswynghels M, Deckers J, De Wolf C, Toussaint W, et al. TCR transgenic clone selection guided by immune receptor analysis and single cell RNA expression of polyclonal responders. BIORXIV. 2024.
- IEEE
- [1]N. Debeuf et al., “TCR transgenic clone selection guided by immune receptor analysis and single cell RNA expression of polyclonal responders,” BIORXIV. 2024.
@misc{01J19P07290S42DTWG26EZTZDB,
abstract = {{Since the precursor frequency of naïve T cells is extremely low, investigating the early steps of antigen-specific T cell activation is challenging. To overcome this detection problem, adoptive transfer of a cohort of T cells purified from T cell receptor (TCR) transgenic donors has been extensively used but is not readily available for emerging pathogens. Constructing TCR transgenic mice from T cell hybridomas is a labor-intensive and sometimes erratic process, since the best clones are selected based on antigen-induced CD69 upregulation or IL-2 production in vitro, and TCR chains are PCR-cloned into expression vectors. Here, we exploited the rapid advances in single cell sequencing and TCR repertoire analysis to select the best clones without hybridoma selection, and generated CORSET8 mice (CORona Spike Epitope specific CD8 T cell), carrying a TCR specific for the Spike protein of SARS-CoV-2. Implementing newly created DALI software for TCR repertoire analysis in single cell analysis enabled the rapid selection of the ideal responder CD8 T cell clone, based on antigen reactivity, proliferation and immunophenotype in vivo. In contrast, a traditional method based on hybridoma technology was unsuccessful. Identified TCR sequences were inserted as synthetic DNA into an expression vector and transgenic CORSET8 donor mice were created. After immunization with Spike/CpG-motifs, mRNA vaccination or SARS-CoV2 infection, CORSET8 T cells strongly proliferated and showed signs of T cell activation. Thus, a combination of TCR repertoire analysis and scRNA immunophenotyping allowed rapid selection of antigen-specific TCR sequences that can be used to generate TCR transgenic mice.}},
author = {{Debeuf, Nincy and Lameire, Sahine and Vanheerswynghels, Manon and Deckers, Julie and De Wolf, Caroline and Toussaint, Wendy and Verbeke, Rein and Verstaen, Kevin and Hammad, Hamida and Vanhee, Stijn and Lambrecht, Bart}},
language = {{eng}},
pages = {{28}},
series = {{BIORXIV}},
title = {{TCR transgenic clone selection guided by immune receptor analysis and single cell RNA expression of polyclonal responders}},
url = {{http://doi.org/10.7554/elife.98344.1}},
year = {{2024}},
}
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