Isolation and enrichment of mouse splenic T cells for ex vivo and in vivo T cell receptor stimulation assays
Summary: Specific antigen recognition by T cell receptor (TCR) activates TCR signaling pathway, leading to T cell proliferation and differentiation into effector and memory cells. Herein, we describe protocols for TCR stimulation assays, including procedures for the isolation and enrichment of mouse...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2021-12-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166721006675 |
| _version_ | 1819174874745143296 |
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| author | Clémence Grosjean Julie Quessada Mathis Nozais Marie Loosveld Dominique Payet-Bornet Cyrille Mionnet |
| author_facet | Clémence Grosjean Julie Quessada Mathis Nozais Marie Loosveld Dominique Payet-Bornet Cyrille Mionnet |
| author_sort | Clémence Grosjean |
| collection | DOAJ |
| description | Summary: Specific antigen recognition by T cell receptor (TCR) activates TCR signaling pathway, leading to T cell proliferation and differentiation into effector and memory cells. Herein, we describe protocols for TCR stimulation assays, including procedures for the isolation and enrichment of mouse splenic T cells for ex vivo TCR stimulation with anti-CD3/CD28 antibodies, and the use of ovalbumin-OT-II mouse model for in vivo TCR stimulation. We applied this protocol to show that MYC protein is essential for T cell proliferation and differentiation.For complete details on the use and execution of this protocol, please refer to Nozais et al. (2021). |
| first_indexed | 2024-12-22T20:45:55Z |
| format | Article |
| id | doaj.art-0d2ce91870624e8f9e0a3dcd01ab8efd |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-22T20:45:55Z |
| publishDate | 2021-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-0d2ce91870624e8f9e0a3dcd01ab8efd2022-12-21T18:13:13ZengElsevierSTAR Protocols2666-16672021-12-0124100961Isolation and enrichment of mouse splenic T cells for ex vivo and in vivo T cell receptor stimulation assaysClémence Grosjean0Julie Quessada1Mathis Nozais2Marie Loosveld3Dominique Payet-Bornet4Cyrille Mionnet5Aix Marseille Univ, CNRS, Inserm, Centre d’immunologie de Marseille-Luminy (CIML), Parc Scientifique de Luminy, Case 906, 13 288 Marseille Cedex 9, France; Corresponding authorAix Marseille Univ, CNRS, Inserm, Centre d’immunologie de Marseille-Luminy (CIML), Parc Scientifique de Luminy, Case 906, 13 288 Marseille Cedex 9, FranceAix Marseille Univ, CNRS, Inserm, Centre d’immunologie de Marseille-Luminy (CIML), Parc Scientifique de Luminy, Case 906, 13 288 Marseille Cedex 9, FranceAix Marseille Univ, CNRS, Inserm, Centre d’immunologie de Marseille-Luminy (CIML), Parc Scientifique de Luminy, Case 906, 13 288 Marseille Cedex 9, France; APHM, Hospital La Timone, Laboratoire d’Hématologie, Marseille, FranceAix Marseille Univ, CNRS, Inserm, Centre d’immunologie de Marseille-Luminy (CIML), Parc Scientifique de Luminy, Case 906, 13 288 Marseille Cedex 9, France; Corresponding authorAix Marseille Univ, CNRS, Inserm, Centre d’immunologie de Marseille-Luminy (CIML), Parc Scientifique de Luminy, Case 906, 13 288 Marseille Cedex 9, France; Corresponding authorSummary: Specific antigen recognition by T cell receptor (TCR) activates TCR signaling pathway, leading to T cell proliferation and differentiation into effector and memory cells. Herein, we describe protocols for TCR stimulation assays, including procedures for the isolation and enrichment of mouse splenic T cells for ex vivo TCR stimulation with anti-CD3/CD28 antibodies, and the use of ovalbumin-OT-II mouse model for in vivo TCR stimulation. We applied this protocol to show that MYC protein is essential for T cell proliferation and differentiation.For complete details on the use and execution of this protocol, please refer to Nozais et al. (2021).http://www.sciencedirect.com/science/article/pii/S2666166721006675Cell BiologyCell cultureCell isolationFlow Cytometry/Mass CytometryCell-based AssaysImmunology |
| spellingShingle | Clémence Grosjean Julie Quessada Mathis Nozais Marie Loosveld Dominique Payet-Bornet Cyrille Mionnet Isolation and enrichment of mouse splenic T cells for ex vivo and in vivo T cell receptor stimulation assays STAR Protocols Cell Biology Cell culture Cell isolation Flow Cytometry/Mass Cytometry Cell-based Assays Immunology |
| title | Isolation and enrichment of mouse splenic T cells for ex vivo and in vivo T cell receptor stimulation assays |
| title_full | Isolation and enrichment of mouse splenic T cells for ex vivo and in vivo T cell receptor stimulation assays |
| title_fullStr | Isolation and enrichment of mouse splenic T cells for ex vivo and in vivo T cell receptor stimulation assays |
| title_full_unstemmed | Isolation and enrichment of mouse splenic T cells for ex vivo and in vivo T cell receptor stimulation assays |
| title_short | Isolation and enrichment of mouse splenic T cells for ex vivo and in vivo T cell receptor stimulation assays |
| title_sort | isolation and enrichment of mouse splenic t cells for ex vivo and in vivo t cell receptor stimulation assays |
| topic | Cell Biology Cell culture Cell isolation Flow Cytometry/Mass Cytometry Cell-based Assays Immunology |
| url | http://www.sciencedirect.com/science/article/pii/S2666166721006675 |
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