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Abstract
This protocol uses the Tg4 Nr4a3-Tocky mouse model to recalibrate T cell activation thresholds and reveals the role that immune checkpoints play in controlling T cell activation. The example approach here uses flow cytometry to characterize quantitative and qualitative changes in splenic CD4+ T cells reactivated in the presence of anti-PD1 immunotherapy. The protocol is optimized for studying anti-PD1 pathway blockade only. The protocol is not compatible with cellular fixation, and T cells should be analyzed immediately after staining.
For complete details on the use and execution of this protocol, please refer to Elliot et al. (2021).
For complete details on the use and execution of this protocol, please refer to Elliot et al. (2021).
Original language | English |
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Article number | 101161 |
Number of pages | 11 |
Journal | STAR Protocols |
Volume | 3 |
Issue number | 1 |
Early online date | 3 Feb 2022 |
DOIs | |
Publication status | Published - 18 Mar 2022 |
Bibliographical note
Funding Information:This work is funded by the MRC ( MR/V009052/1 ). The graphical abstract and Figure 1 were created using https://biorender.com .
Publisher Copyright:
© 2022 The Author(s)
Keywords
- Cancer
- Cell Biology
- Flow Cytometry/Mass Cytometry
- Immunology
- Model Organisms
ASJC Scopus subject areas
- General Biochemistry,Genetics and Molecular Biology
- General Neuroscience
- General Immunology and Microbiology
- General Medicine
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Negative feedback control of T cells in tolerance and cancer - from pathways to biomarkers
Bending, D. (Principal Investigator)
1/02/21 → 31/01/26
Project: Research Councils