TCR transgenic clone selection guided by immune receptor analysis and single-cell RNA expression of polyclonal responders
Since the precursor frequency of naive 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...
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eLife Sciences Publications Ltd
2024-12-01
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| Online Access: | https://elifesciences.org/articles/98344 |
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| author | Nincy Debeuf Sahine Lameire Manon Vanheerswynghels Julie Deckers Caroline De Wolf Wendy Toussaint Rein Verbeke Kevin Verstaen Hamida Hammad Stijn Vanhee Bart N Lambrecht |
| author_facet | Nincy Debeuf Sahine Lameire Manon Vanheerswynghels Julie Deckers Caroline De Wolf Wendy Toussaint Rein Verbeke Kevin Verstaen Hamida Hammad Stijn Vanhee Bart N Lambrecht |
| author_sort | Nincy Debeuf |
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| description | Since the precursor frequency of naive 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 polymerase chain reaction (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. 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-CoV-2 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. |
| format | Article |
| id | doaj-art-b5e1f9ec55a546aa99c0a96aacadfd5e |
| institution | Kabale University |
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| language | English |
| publishDate | 2024-12-01 |
| publisher | eLife Sciences Publications Ltd |
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| series | eLife |
| spelling | doaj-art-b5e1f9ec55a546aa99c0a96aacadfd5e2025-01-27T10:17:39ZengeLife Sciences Publications LtdeLife2050-084X2024-12-011310.7554/eLife.98344TCR transgenic clone selection guided by immune receptor analysis and single-cell RNA expression of polyclonal respondersNincy Debeuf0https://orcid.org/0000-0002-4754-4772Sahine Lameire1Manon Vanheerswynghels2Julie Deckers3Caroline De Wolf4Wendy Toussaint5Rein Verbeke6Kevin Verstaen7Hamida Hammad8Stijn Vanhee9Bart N Lambrecht10https://orcid.org/0000-0003-4376-6834Laboratory of Immunoregulation and Mucosal Immunology, VIB Center for Inflammation Research, Ghent, Belgium; Department of Internal Medicine and Pediatrics, Ghent University, Ghent, BelgiumLaboratory of Immunoregulation and Mucosal Immunology, VIB Center for Inflammation Research, Ghent, Belgium; Department of Internal Medicine and Pediatrics, Ghent University, Ghent, BelgiumLaboratory of Immunoregulation and Mucosal Immunology, VIB Center for Inflammation Research, Ghent, Belgium; Department of Internal Medicine and Pediatrics, Ghent University, Ghent, BelgiumLaboratory of Immunoregulation and Mucosal Immunology, VIB Center for Inflammation Research, Ghent, Belgium; Department of Internal Medicine and Pediatrics, Ghent University, Ghent, BelgiumLaboratory of Immunoregulation and Mucosal Immunology, VIB Center for Inflammation Research, Ghent, Belgium; Department of Internal Medicine and Pediatrics, Ghent University, Ghent, BelgiumLaboratory of Immunoregulation and Mucosal Immunology, VIB Center for Inflammation Research, Ghent, Belgium; Department of Internal Medicine and Pediatrics, Ghent University, Ghent, BelgiumLaboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmaceutical Sciences, Ghent University, Ghent, BelgiumVIB Single Cell Core, VIB Center, Ghent, Belgium; Department of Applied Mathematics, Computer Science and Statistics, Ghent University, Ghent, BelgiumLaboratory of Immunoregulation and Mucosal Immunology, VIB Center for Inflammation Research, Ghent, Belgium; Department of Internal Medicine and Pediatrics, Ghent University, Ghent, BelgiumLaboratory of Immunoregulation and Mucosal Immunology, VIB Center for Inflammation Research, Ghent, Belgium; Department of Internal Medicine and Pediatrics, Ghent University, Ghent, Belgium; Department of Head and Skin, Ghent University, Ghent, BelgiumLaboratory of Immunoregulation and Mucosal Immunology, VIB Center for Inflammation Research, Ghent, Belgium; Department of Internal Medicine and Pediatrics, Ghent University, Ghent, Belgium; Department of Pulmonary Medicine, Erasmus University Medical Center Rotterdam, Rotterdam, NetherlandsSince the precursor frequency of naive 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 polymerase chain reaction (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. 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-CoV-2 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.https://elifesciences.org/articles/98344SARS-CoV-2TCR transgenic mousesingle-cell RNA analysisVDJseq |
| spellingShingle | Nincy Debeuf Sahine Lameire Manon Vanheerswynghels Julie Deckers Caroline De Wolf Wendy Toussaint Rein Verbeke Kevin Verstaen Hamida Hammad Stijn Vanhee Bart N Lambrecht TCR transgenic clone selection guided by immune receptor analysis and single-cell RNA expression of polyclonal responders eLife SARS-CoV-2 TCR transgenic mouse single-cell RNA analysis VDJseq |
| title | TCR transgenic clone selection guided by immune receptor analysis and single-cell RNA expression of polyclonal responders |
| title_full | TCR transgenic clone selection guided by immune receptor analysis and single-cell RNA expression of polyclonal responders |
| title_fullStr | TCR transgenic clone selection guided by immune receptor analysis and single-cell RNA expression of polyclonal responders |
| title_full_unstemmed | TCR transgenic clone selection guided by immune receptor analysis and single-cell RNA expression of polyclonal responders |
| title_short | TCR transgenic clone selection guided by immune receptor analysis and single-cell RNA expression of polyclonal responders |
| title_sort | tcr transgenic clone selection guided by immune receptor analysis and single cell rna expression of polyclonal responders |
| topic | SARS-CoV-2 TCR transgenic mouse single-cell RNA analysis VDJseq |
| url | https://elifesciences.org/articles/98344 |
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