Separate signaling events control TCR downregulation and T cell activation in primary human T cells
Abstract Introduction T‐cell antigen receptor (TCR) interaction with cognate peptide:MHC complexes trigger clustering of TCR:CD3 complexes and signal transduction. Triggered TCR:CD3 complexes are rapidly internalized and degraded in a process called ligand‐induced TCR downregulation. Classic studies...
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| Format: | Article |
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Wiley
2021-03-01
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| Series: | Immunity, Inflammation and Disease |
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| Online Access: | https://doi.org/10.1002/iid3.383 |
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| author | Lieve E. H. van derDonk Louis S. Ates Jet van derSpek Laura M. Tukker Teunis B. H. Geijtenbeek Jeroen W. J. vanHeijst |
| author_facet | Lieve E. H. van derDonk Louis S. Ates Jet van derSpek Laura M. Tukker Teunis B. H. Geijtenbeek Jeroen W. J. vanHeijst |
| author_sort | Lieve E. H. van derDonk |
| collection | DOAJ |
| description | Abstract Introduction T‐cell antigen receptor (TCR) interaction with cognate peptide:MHC complexes trigger clustering of TCR:CD3 complexes and signal transduction. Triggered TCR:CD3 complexes are rapidly internalized and degraded in a process called ligand‐induced TCR downregulation. Classic studies in immortalized T‐cell lines have revealed a major role for the Src family kinase Lck in TCR downregulation. However, to what extent a similar mechanism operates in primary human T cells remains unclear. Methods Here, we developed an anti‐CD3‐mediated TCR downregulation assay, in which T‐cell gene expression in primary human T cells can be knocked down by microRNA constructs. In parallel, we used CRISPR/Cas9‐mediated knockout in Jurkat cells for validation experiments. Results We efficiently knocked down the expression of tyrosine kinases Lck, Fyn, and ZAP70, and found that, whereas this impaired T cell activation and effector function, TCR downregulation was not affected. Although TCR downregulation was marginally inhibited by the simultaneous knockdown of Lck and Fyn, its full abrogation required broad‐acting tyrosine kinase inhibitors. Conclusions These data suggest that there is substantial redundancy in the contribution of individual tyrosine kinases to TCR downregulation in primary human T cells. Our results highlight that TCR downregulation and T cell activation are controlled by different signaling events and illustrate the need for further research to untangle these processes. |
| format | Article |
| id | doaj-art-53079b524d644ffd827d286d6ed39ed9 |
| institution | DOAJ |
| issn | 2050-4527 |
| language | English |
| publishDate | 2021-03-01 |
| publisher | Wiley |
| record_format | Article |
| series | Immunity, Inflammation and Disease |
| spelling | doaj-art-53079b524d644ffd827d286d6ed39ed92025-08-20T03:08:21ZengWileyImmunity, Inflammation and Disease2050-45272021-03-019122323810.1002/iid3.383Separate signaling events control TCR downregulation and T cell activation in primary human T cellsLieve E. H. van derDonk0Louis S. Ates1Jet van derSpek2Laura M. Tukker3Teunis B. H. Geijtenbeek4Jeroen W. J. vanHeijst5Department of Experimental Immunology, Amsterdam Infection and Immunity Institute, Amsterdam UMC University of Amsterdam Amsterdam The NetherlandsDepartment of Experimental Immunology, Amsterdam Infection and Immunity Institute, Amsterdam UMC University of Amsterdam Amsterdam The NetherlandsDepartment of Experimental Immunology, Amsterdam Infection and Immunity Institute, Amsterdam UMC University of Amsterdam Amsterdam The NetherlandsDepartment of Experimental Immunology, Amsterdam Infection and Immunity Institute, Amsterdam UMC University of Amsterdam Amsterdam The NetherlandsDepartment of Experimental Immunology, Amsterdam Infection and Immunity Institute, Amsterdam UMC University of Amsterdam Amsterdam The NetherlandsDepartment of Experimental Immunology, Amsterdam Infection and Immunity Institute, Amsterdam UMC University of Amsterdam Amsterdam The NetherlandsAbstract Introduction T‐cell antigen receptor (TCR) interaction with cognate peptide:MHC complexes trigger clustering of TCR:CD3 complexes and signal transduction. Triggered TCR:CD3 complexes are rapidly internalized and degraded in a process called ligand‐induced TCR downregulation. Classic studies in immortalized T‐cell lines have revealed a major role for the Src family kinase Lck in TCR downregulation. However, to what extent a similar mechanism operates in primary human T cells remains unclear. Methods Here, we developed an anti‐CD3‐mediated TCR downregulation assay, in which T‐cell gene expression in primary human T cells can be knocked down by microRNA constructs. In parallel, we used CRISPR/Cas9‐mediated knockout in Jurkat cells for validation experiments. Results We efficiently knocked down the expression of tyrosine kinases Lck, Fyn, and ZAP70, and found that, whereas this impaired T cell activation and effector function, TCR downregulation was not affected. Although TCR downregulation was marginally inhibited by the simultaneous knockdown of Lck and Fyn, its full abrogation required broad‐acting tyrosine kinase inhibitors. Conclusions These data suggest that there is substantial redundancy in the contribution of individual tyrosine kinases to TCR downregulation in primary human T cells. Our results highlight that TCR downregulation and T cell activation are controlled by different signaling events and illustrate the need for further research to untangle these processes.https://doi.org/10.1002/iid3.383CD4+ T cellsprotein tyrosine kinasesT‐cell activation | T‐cell receptorTCR downregulation |
| spellingShingle | Lieve E. H. van derDonk Louis S. Ates Jet van derSpek Laura M. Tukker Teunis B. H. Geijtenbeek Jeroen W. J. vanHeijst Separate signaling events control TCR downregulation and T cell activation in primary human T cells Immunity, Inflammation and Disease CD4+ T cells protein tyrosine kinases T‐cell activation | T‐cell receptor TCR downregulation |
| title | Separate signaling events control TCR downregulation and T cell activation in primary human T cells |
| title_full | Separate signaling events control TCR downregulation and T cell activation in primary human T cells |
| title_fullStr | Separate signaling events control TCR downregulation and T cell activation in primary human T cells |
| title_full_unstemmed | Separate signaling events control TCR downregulation and T cell activation in primary human T cells |
| title_short | Separate signaling events control TCR downregulation and T cell activation in primary human T cells |
| title_sort | separate signaling events control tcr downregulation and t cell activation in primary human t cells |
| topic | CD4+ T cells protein tyrosine kinases T‐cell activation | T‐cell receptor TCR downregulation |
| url | https://doi.org/10.1002/iid3.383 |
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