The role of the different CD3γ domains in TCR expression and signaling
The CD3 subunits of the T-cell antigen receptor (TCR) play a central role in regulation of surface TCR expression levels. Humans who lack CD3γ (γ—) show reduced surface TCR expression levels and abolished phorbol ester (PMA)-induced TCR down-regulation. The response to PMA is mediated by a double le...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-09-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2022.978658/full |
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| author | Beatriz Garcillán Rebeca F. Megino Marta Herrero-Alonso Alberto C. Guardo Veronica Perez-Flores Claudia Juraske Claudia Juraske Claudia Juraske Claudia Juraske Vincent Idstein Vincent Idstein Vincent Idstein Vincent Idstein Jose M. Martin-Fernandez Carsten Geisler Wolfgang W. A. Schamel Wolfgang W. A. Schamel Wolfgang W. A. Schamel Ana V. Marin Jose R. Regueiro |
| author_facet | Beatriz Garcillán Rebeca F. Megino Marta Herrero-Alonso Alberto C. Guardo Veronica Perez-Flores Claudia Juraske Claudia Juraske Claudia Juraske Claudia Juraske Vincent Idstein Vincent Idstein Vincent Idstein Vincent Idstein Jose M. Martin-Fernandez Carsten Geisler Wolfgang W. A. Schamel Wolfgang W. A. Schamel Wolfgang W. A. Schamel Ana V. Marin Jose R. Regueiro |
| author_sort | Beatriz Garcillán |
| collection | DOAJ |
| description | The CD3 subunits of the T-cell antigen receptor (TCR) play a central role in regulation of surface TCR expression levels. Humans who lack CD3γ (γ—) show reduced surface TCR expression levels and abolished phorbol ester (PMA)-induced TCR down-regulation. The response to PMA is mediated by a double leucine motif in the intracellular (IC) domain of CD3γ. However, the molecular cause of the reduced TCR surface expression in γ— lymphocytes is still not known. We used retroviral vectors carrying wild type CD3γ or CD3δ or the following chimeras (EC-extracellular, TM-transmembrane and IC): δECγTMγIC (δγγ for short), γγδ, γδδ and γγ-. Expression of γγγ, γγδ, γδδ or γγ- in the γ— T cell line JGN, which lacks surface TCR, demonstrated that cell surface TCR levels in JGN were dependent on the EC domain of CD3γ and could not be replaced by the one of CD3δ. In JGN and primary γ— patient T cells, the tested chimeras confirmed that the response to PMA maps to the IC domain of CD3γ. Since protein homology explains these results better than domain structure, we conclude that CD3γ contributes conformational cues that improve surface TCR expression, likely at the assembly or membrane transport steps. In JGN cells all chimeric TCRs were signalling competent. However, an IC domain at CD3γ was required for TCR-induced IL-2 and TNF-α production and CD69 expression, indicating that a TCR without a CD3γ IC domain has altered signalling capabilities. |
| first_indexed | 2024-04-11T14:05:21Z |
| format | Article |
| id | doaj.art-ee3075c5866d4f45b4af9db868d1fc27 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-04-11T14:05:21Z |
| publishDate | 2022-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-ee3075c5866d4f45b4af9db868d1fc272022-12-22T04:19:56ZengFrontiers Media S.A.Frontiers in Immunology1664-32242022-09-011310.3389/fimmu.2022.978658978658The role of the different CD3γ domains in TCR expression and signalingBeatriz Garcillán0Rebeca F. Megino1Marta Herrero-Alonso2Alberto C. Guardo3Veronica Perez-Flores4Claudia Juraske5Claudia Juraske6Claudia Juraske7Claudia Juraske8Vincent Idstein9Vincent Idstein10Vincent Idstein11Vincent Idstein12Jose M. Martin-Fernandez13Carsten Geisler14Wolfgang W. A. Schamel15Wolfgang W. A. Schamel16Wolfgang W. A. Schamel17Ana V. Marin18Jose R. Regueiro19Department of Immunology, Ophthalmology and Ear, Nose and Throat (ENT), Complutense University School of Medicine and 12 de Octubre Health Research Institute (imas12), Madrid, SpainDepartment of Immunology, Ophthalmology and Ear, Nose and Throat (ENT), Complutense University School of Medicine and 12 de Octubre Health Research Institute (imas12), Madrid, SpainDepartment of Immunology, Ophthalmology and Ear, Nose and Throat (ENT), Complutense University School of Medicine and 12 de Octubre Health Research Institute (imas12), Madrid, SpainDepartment of Immunology, Ophthalmology and Ear, Nose and Throat (ENT), Complutense University School of Medicine and 12 de Octubre Health Research Institute (imas12), Madrid, SpainDepartment of Immunology, Ophthalmology and Ear, Nose and Throat (ENT), Complutense University School of Medicine and 12 de Octubre Health Research Institute (imas12), Madrid, SpainSignaling Research Centers BIOSS and CIBSS, University of Freiburg, Freiburg, GermanyDepartment of Immunology, Faculty of Biology, University of Freiburg, Freiburg, GermanyCentre for Chronic Immunodeficiency (CCI), Medical Center Freiburg and Faculty of Medicine, University of Freiburg, Freiburg, GermanySpemann Graduate School of Biology and Medicine (SGBM), University of Freiburg, Freiburg, GermanySignaling Research Centers BIOSS and CIBSS, University of Freiburg, Freiburg, GermanyDepartment of Immunology, Faculty of Biology, University of Freiburg, Freiburg, GermanyCentre for Chronic Immunodeficiency (CCI), Medical Center Freiburg and Faculty of Medicine, University of Freiburg, Freiburg, GermanySpemann Graduate School of Biology and Medicine (SGBM), University of Freiburg, Freiburg, GermanyDepartment of Immunology, Ophthalmology and Ear, Nose and Throat (ENT), Complutense University School of Medicine and 12 de Octubre Health Research Institute (imas12), Madrid, SpainThe LEO Foundation Skin Immunology Research Center, Department of Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, DenmarkSignaling Research Centers BIOSS and CIBSS, University of Freiburg, Freiburg, GermanyDepartment of Immunology, Faculty of Biology, University of Freiburg, Freiburg, GermanyCentre for Chronic Immunodeficiency (CCI), Medical Center Freiburg and Faculty of Medicine, University of Freiburg, Freiburg, GermanyDepartment of Immunology, Ophthalmology and Ear, Nose and Throat (ENT), Complutense University School of Medicine and 12 de Octubre Health Research Institute (imas12), Madrid, SpainDepartment of Immunology, Ophthalmology and Ear, Nose and Throat (ENT), Complutense University School of Medicine and 12 de Octubre Health Research Institute (imas12), Madrid, SpainThe CD3 subunits of the T-cell antigen receptor (TCR) play a central role in regulation of surface TCR expression levels. Humans who lack CD3γ (γ—) show reduced surface TCR expression levels and abolished phorbol ester (PMA)-induced TCR down-regulation. The response to PMA is mediated by a double leucine motif in the intracellular (IC) domain of CD3γ. However, the molecular cause of the reduced TCR surface expression in γ— lymphocytes is still not known. We used retroviral vectors carrying wild type CD3γ or CD3δ or the following chimeras (EC-extracellular, TM-transmembrane and IC): δECγTMγIC (δγγ for short), γγδ, γδδ and γγ-. Expression of γγγ, γγδ, γδδ or γγ- in the γ— T cell line JGN, which lacks surface TCR, demonstrated that cell surface TCR levels in JGN were dependent on the EC domain of CD3γ and could not be replaced by the one of CD3δ. In JGN and primary γ— patient T cells, the tested chimeras confirmed that the response to PMA maps to the IC domain of CD3γ. Since protein homology explains these results better than domain structure, we conclude that CD3γ contributes conformational cues that improve surface TCR expression, likely at the assembly or membrane transport steps. In JGN cells all chimeric TCRs were signalling competent. However, an IC domain at CD3γ was required for TCR-induced IL-2 and TNF-α production and CD69 expression, indicating that a TCR without a CD3γ IC domain has altered signalling capabilities.https://www.frontiersin.org/articles/10.3389/fimmu.2022.978658/fullCD3 chimerasT cell receptorCD3δCD3γdomains |
| spellingShingle | Beatriz Garcillán Rebeca F. Megino Marta Herrero-Alonso Alberto C. Guardo Veronica Perez-Flores Claudia Juraske Claudia Juraske Claudia Juraske Claudia Juraske Vincent Idstein Vincent Idstein Vincent Idstein Vincent Idstein Jose M. Martin-Fernandez Carsten Geisler Wolfgang W. A. Schamel Wolfgang W. A. Schamel Wolfgang W. A. Schamel Ana V. Marin Jose R. Regueiro The role of the different CD3γ domains in TCR expression and signaling Frontiers in Immunology CD3 chimeras T cell receptor CD3δ CD3γ domains |
| title | The role of the different CD3γ domains in TCR expression and signaling |
| title_full | The role of the different CD3γ domains in TCR expression and signaling |
| title_fullStr | The role of the different CD3γ domains in TCR expression and signaling |
| title_full_unstemmed | The role of the different CD3γ domains in TCR expression and signaling |
| title_short | The role of the different CD3γ domains in TCR expression and signaling |
| title_sort | role of the different cd3γ domains in tcr expression and signaling |
| topic | CD3 chimeras T cell receptor CD3δ CD3γ domains |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2022.978658/full |
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