CD3G or CD3D Knockdown in Mature, but Not Immature, T Lymphocytes Similarly Cripples the Human TCRαβ Complex
The human αβ T-cell receptor (TCR) is composed of a variable heterodimer (TCRαβ) and three invariant dimers (CD3γε, CD3δε, and ζζ/CD247). The role of each invariant chain in the stepwise interactions among TCR chains along the assembly is still not fully understood. Despite the high sequence homolog...
| Autores: | , , , , , , , , , , , , |
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| Tipo de recurso: | artículo |
| Estado: | Versión publicada |
| Fecha de publicación: | 2021 |
| País: | España |
| Institución: | Consejo Superior de Investigaciones Científicas (CSIC) |
| Repositorio: | DIGITAL.CSIC. Repositorio Institucional del CSIC |
| OAI Identifier: | oai:digital.csic.es:10261/263310 |
| Acceso en línea: | http://hdl.handle.net/10261/263310 |
| Access Level: | acceso abierto |
| Palabra clave: | CD3G CD3D shRNA knockdown T-cell receptor (TCR) TCR assembly T-cell progenitors Immunodeficiency |
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CD3G or CD3D Knockdown in Mature, but Not Immature, T Lymphocytes Similarly Cripples the Human TCRαβ ComplexGarcillán, BeatrizFuentes, PatriciaMarín, Ana V.Megino, Rebeca F.Chacon-Arguedas, DanielMazariegos, Marina S.Jiménez-Reinoso, AnaïsMuñoz-Ruiz, MiguelLaborda, Raquel G.Cárdenas, Paula P.Fernández-Malavé, EdgarToribio, María LuisaRegueiro, José R.CD3GCD3DshRNA knockdownT-cell receptor (TCR)TCR assemblyT-cell progenitorsImmunodeficiencyThe human αβ T-cell receptor (TCR) is composed of a variable heterodimer (TCRαβ) and three invariant dimers (CD3γε, CD3δε, and ζζ/CD247). The role of each invariant chain in the stepwise interactions among TCR chains along the assembly is still not fully understood. Despite the high sequence homology between CD3γ and CD3δ, the clinical consequences of the corresponding immunodeficiencies (ID) in humans are very different (mild and severe, respectively), and mouse models do not recapitulate findings in human ID. To try to understand such disparities, we stably knocked down (KD) CD3D or CD3G expression in the human Jurkat T-cell line and analyzed comparatively their impact on TCRαβ assembly, transport, and surface expression. The results indicated that TCR ensembles were less stable and CD3ε levels were lower when CD3γ, rather than CD3δ, was scarce. However, both defective TCR ensembles were strongly retained in the ER, lacked ζζ/CD247, and barely reached the T-cell surface (<11% of normal controls) in any of the CD3 KD cells. This is in sharp contrast to human CD3γ ID, whose mature T cells express higher levels of surface TCR (>30% vs. normal controls). CD3 KD of human T-cell progenitors followed by mouse fetal thymus organ cultures showed high plasticity in emerging immature polyclonal T lymphocytes that allowed for the expression of significant TCR levels which may then signal for survival in CD3γ, but not in CD3δ deficiency, and explain the immunological and clinical disparities of such ID cases.Ministerio de Economía y Competitividad (MINECO RTI2018-095673-B-I00 and PID2019-105623RB-I00), Comunidad Autónoma deMadrid (CAM B2017/BMD3673), and Asociación Española Contra el Cáncer (AECC PROYE20084REGU and CICPF18030TORI), Fundación Ramón Areces, and Fundación Unoentrecienmil. AM was supported by Complutense/Harvard University scholarship CT46/15 and AJ-R by MINECO scholarship BES-2012-055054Ministerio de Economía y Competitividad (España)Comunidad de MadridAsociación Española Contra el CáncerFundación Ramón ArecesFundación UnoentrecienmilConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]2022202220212022info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/263310reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Ingléshttp://dx.doi.org/10.3389/fcell.2021.608490Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2633102026-05-22T06:33:51Z |
| dc.title.none.fl_str_mv |
CD3G or CD3D Knockdown in Mature, but Not Immature, T Lymphocytes Similarly Cripples the Human TCRαβ Complex |
| title |
CD3G or CD3D Knockdown in Mature, but Not Immature, T Lymphocytes Similarly Cripples the Human TCRαβ Complex |
| spellingShingle |
CD3G or CD3D Knockdown in Mature, but Not Immature, T Lymphocytes Similarly Cripples the Human TCRαβ Complex Garcillán, Beatriz CD3G CD3D shRNA knockdown T-cell receptor (TCR) TCR assembly T-cell progenitors Immunodeficiency |
| title_short |
CD3G or CD3D Knockdown in Mature, but Not Immature, T Lymphocytes Similarly Cripples the Human TCRαβ Complex |
| title_full |
CD3G or CD3D Knockdown in Mature, but Not Immature, T Lymphocytes Similarly Cripples the Human TCRαβ Complex |
| title_fullStr |
CD3G or CD3D Knockdown in Mature, but Not Immature, T Lymphocytes Similarly Cripples the Human TCRαβ Complex |
| title_full_unstemmed |
CD3G or CD3D Knockdown in Mature, but Not Immature, T Lymphocytes Similarly Cripples the Human TCRαβ Complex |
| title_sort |
CD3G or CD3D Knockdown in Mature, but Not Immature, T Lymphocytes Similarly Cripples the Human TCRαβ Complex |
| dc.creator.none.fl_str_mv |
Garcillán, Beatriz Fuentes, Patricia Marín, Ana V. Megino, Rebeca F. Chacon-Arguedas, Daniel Mazariegos, Marina S. Jiménez-Reinoso, Anaïs Muñoz-Ruiz, Miguel Laborda, Raquel G. Cárdenas, Paula P. Fernández-Malavé, Edgar Toribio, María Luisa Regueiro, José R. |
| author |
Garcillán, Beatriz |
| author_facet |
Garcillán, Beatriz Fuentes, Patricia Marín, Ana V. Megino, Rebeca F. Chacon-Arguedas, Daniel Mazariegos, Marina S. Jiménez-Reinoso, Anaïs Muñoz-Ruiz, Miguel Laborda, Raquel G. Cárdenas, Paula P. Fernández-Malavé, Edgar Toribio, María Luisa Regueiro, José R. |
| author_role |
author |
| author2 |
Fuentes, Patricia Marín, Ana V. Megino, Rebeca F. Chacon-Arguedas, Daniel Mazariegos, Marina S. Jiménez-Reinoso, Anaïs Muñoz-Ruiz, Miguel Laborda, Raquel G. Cárdenas, Paula P. Fernández-Malavé, Edgar Toribio, María Luisa Regueiro, José R. |
| author2_role |
author author author author author author author author author author author author |
| dc.contributor.none.fl_str_mv |
Ministerio de Economía y Competitividad (España) Comunidad de Madrid Asociación Española Contra el Cáncer Fundación Ramón Areces Fundación Unoentrecienmil Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72] |
| dc.subject.none.fl_str_mv |
CD3G CD3D shRNA knockdown T-cell receptor (TCR) TCR assembly T-cell progenitors Immunodeficiency |
| topic |
CD3G CD3D shRNA knockdown T-cell receptor (TCR) TCR assembly T-cell progenitors Immunodeficiency |
| description |
The human αβ T-cell receptor (TCR) is composed of a variable heterodimer (TCRαβ) and three invariant dimers (CD3γε, CD3δε, and ζζ/CD247). The role of each invariant chain in the stepwise interactions among TCR chains along the assembly is still not fully understood. Despite the high sequence homology between CD3γ and CD3δ, the clinical consequences of the corresponding immunodeficiencies (ID) in humans are very different (mild and severe, respectively), and mouse models do not recapitulate findings in human ID. To try to understand such disparities, we stably knocked down (KD) CD3D or CD3G expression in the human Jurkat T-cell line and analyzed comparatively their impact on TCRαβ assembly, transport, and surface expression. The results indicated that TCR ensembles were less stable and CD3ε levels were lower when CD3γ, rather than CD3δ, was scarce. However, both defective TCR ensembles were strongly retained in the ER, lacked ζζ/CD247, and barely reached the T-cell surface (<11% of normal controls) in any of the CD3 KD cells. This is in sharp contrast to human CD3γ ID, whose mature T cells express higher levels of surface TCR (>30% vs. normal controls). CD3 KD of human T-cell progenitors followed by mouse fetal thymus organ cultures showed high plasticity in emerging immature polyclonal T lymphocytes that allowed for the expression of significant TCR levels which may then signal for survival in CD3γ, but not in CD3δ deficiency, and explain the immunological and clinical disparities of such ID cases. |
| publishDate |
2021 |
| dc.date.none.fl_str_mv |
2021 2022 2022 2022 |
| dc.type.none.fl_str_mv |
info:eu-repo/semantics/article http://purl.org/coar/resource_type/c_6501 Publisher's version info:eu-repo/semantics/publishedVersion |
| format |
article |
| status_str |
publishedVersion |
| dc.identifier.none.fl_str_mv |
http://hdl.handle.net/10261/263310 |
| url |
http://hdl.handle.net/10261/263310 |
| dc.language.none.fl_str_mv |
Inglés |
| language_invalid_str_mv |
Inglés |
| dc.relation.none.fl_str_mv |
http://dx.doi.org/10.3389/fcell.2021.608490 Sí |
| dc.rights.none.fl_str_mv |
info:eu-repo/semantics/openAccess |
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openAccess |
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reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC instname:Consejo Superior de Investigaciones Científicas (CSIC) |
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Consejo Superior de Investigaciones Científicas (CSIC) |
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DIGITAL.CSIC. Repositorio Institucional del CSIC |
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DIGITAL.CSIC. Repositorio Institucional del CSIC |
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15,81155 |