Analyse intégrative de l’activation des lymphocytes T

Publications de l’équipe

Année de publication : 2020

Chabaud M1,2,3, Paillon N1, Gaus K2,3, Hivroz C1. (2020 Apr 2)

Mechanobiology of antigen-induced T cell arrest.

Biology of the Cell : DOI : 10.1111/boc.201900093 En savoir plus
Résumé

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Année de publication : 2019

Gehrmann U1,2, Burbage M3, Zueva E3, Goudot C3, Esnault C4, Ye M3, Carpier JM3, Burgdorf N3, Hoyler T3, Suarez G3, Joannas L3, Heurtebise-Chrétien S3, Durand S5,6, Panes R7,8, Bellemare-Pelletier A8, Sáez PJ3, Aprahamian F5,6, Lefevre D5,6, Adoue V9, Zine El Aabidine A4, Muhammad Ahmad M4, Hivroz C3, Joffre O10, Cammas F11,12, Kroemer G5,6,13,14,15, Gagnon E7,8, Andrau JC4, Amigorena S1. (2019 Dec 17)

Critical role for TRIM28 and HP1β/γ in the epigenetic control of T cell metabolic reprograming and effector differentiation.

Proceedings of the National Academy of Sciences : 116 : Proc Natl Acad Sci U S A. 2019 Dec 17;116(51):25839-25849. doi: 10.1073/pnas.1901639116. Epub 2019 Nov 27. : 25839,25849 : DOI : 10.1073/pnas.1901639116 En savoir plus
Résumé

Naive CD4+ T lymphocytes differentiate into different effector types, including helper and regulatory cells (Th and Treg, respectively). Heritable gene expression programs that define these effector types are established during differentiation, but little is known about the epigenetic mechanisms that install and maintain these programs. Here, we use mice defective for different components of heterochromatin-dependent gene silencing to investigate the epigenetic control of CD4+ T cell plasticity. We show that, upon T cell receptor (TCR) engagement, naive and regulatory T cells defective for TRIM28 (an epigenetic adaptor for histone binding modules) or for heterochromatin protein 1 β and γ isoforms (HP1β/γ, 2 histone-binding factors involved in gene silencing) fail to effectively signal through the PI3K-AKT-mTOR axis and switch to glycolysis. While differentiation of naive TRIM28-/- T cells into cytokine-producing effector T cells is impaired, resulting in reduced induction of autoimmune colitis, TRIM28-/- regulatory T cells also fail to expand in vivo and to suppress autoimmunity effectively. Using a combination of transcriptome and chromatin immunoprecipitation-sequencing (ChIP-seq) analyses for H3K9me3, H3K9Ac, and RNA polymerase II, we show that reduced effector differentiation correlates with impaired transcriptional silencing at distal regulatory regions of a defined set of Treg-associated genes, including, for example, NRP1 or Snai3. We conclude that TRIM28 and HP1β/γ control metabolic reprograming through epigenetic silencing of a defined set of Treg-characteristic genes, thus allowing effective T cell expansion and differentiation into helper and regulatory phenotypes.

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Saveanu L1,2,3,4, Zucchetti AE5, Evnouchidou I1,2,3,4,6, Ardouin L5, Hivroz C5. (2019 Sep 2)

Is there a place and role for endocytic TCR signaling?

Immunological reviews : 291(1) : 57-74 : DOI : 10.1111/imr.12764 En savoir plus
Résumé

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Loredana Saveanu, Andres E Zucchetti, Irini Evnouchidou, Laurence Ardouin, Claire Hivroz (2019 Aug 13)

Is there a place and role for endocytic TCR signaling?

Immunological reviews : 57-74 : DOI : 10.1111/imr.12764 En savoir plus
Résumé

T-lymphocyte activation relies on the cognate recognition by the TCR of the MHC-associated peptide ligand (pMHC) presented at the surface of an antigen-presenting cell (APC). This leads to the dynamic formation of a cognate contact between the T lymphocyte and the APC: the immune synapse (IS). Engagement of the TCR by the pMHC in the synaptic zone induces a cascade of signaling events leading to phosphorylation and dephosphorylation of proteins and lipids, which ultimately shapes the response of T lymphocytes. Although the engagement of the T-cell receptor (TCR) takes place at the plasma membrane, the TCR/CD3 complexes and the signaling molecules involved in transduction of the TCR signal are also present in intracellular membrane pools. These pools, which are both endocytic and exocytic, have tentatively been characterized by several groups including ours. We will herein summarize what is known on the intracellular pools of TCR signaling components. We will discuss their origin and the mechanisms involved in their mobility at the IS. Finally, we will propose several hypotheses concerning the functional role(s) that these intracellular pools might play in T-cell activation. We will also discuss the tools that could be used to test these hypotheses.

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Andres Ernesto Zucchetti, Laurence Bataille, Jean-Marie Carpier, Stéphanie Dogniaux, Mabel San Roman-Jouve, Mathieu Maurin, Michael W Stuck, Rosa M Rios, Cosima T Baldari, Gregory J Pazour, Claire Hivroz (2019 Jun 30)

Tethering of vesicles to the Golgi by GMAP210 controls LAT delivery to the immune synapse.

Nature communications : 2864 : DOI : 10.1038/s41467-019-10891-w En savoir plus
Résumé

The T cell immune synapse is a site of intense vesicular trafficking. Here we show that the golgin GMAP210, known to capture vesicles and organize membrane traffic at the Golgi, is involved in the vesicular transport of LAT to the immune synapse. Upon activation, more GMAP210 interact with LAT-containing vesicles and go together with LAT to the immune synapse. Regulating LAT recruitment and LAT-dependent signaling, GMAP210 controls T cell activation. Using a rerouting and capture assay, we show that GMAP210 captures VAMP7-decorated vesicles. Overexpressing different domains of GMAP210, we also show that GMAP210 allows their specific delivery to the immune synapse by tethering LAT-vesicles to the Golgi. Finally, in a model of ectopic expression of LAT in ciliated cells, we show that GMAP210 tethering activity controls the delivery of LAT to the cilium. Hence, our results reveal a function for the golgin GMAP210 conveying specific vesicles to the immune synapse.

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Année de publication : 2018

Jean-Marie Carpier, Andres E Zucchetti, Laurence Bataille, Stéphanie Dogniaux, Massiullah Shafaq-Zadah, Sabine Bardin, Marco Lucchino, Mathieu Maurin, Leonel D Joannas, Joao Gamelas Magalhaes, Ludger Johannes, Thierry Galli, Bruno Goud, Claire Hivroz (2018 Feb 15)

Rab6-dependent retrograde traffic of LAT controls immune synapse formation and T cell activation.

The Journal of experimental medicine : 1245-1265 : DOI : 10.1084/jem.20162042 En savoir plus
Résumé

The adapter molecule linker for activation of T cells (LAT) orchestrates the formation of signalosomes upon T cell receptor (TCR) stimulation. LAT is present in different intracellular pools and is dynamically recruited to the immune synapse upon stimulation. However, the intracellular traffic of LAT and its function in T lymphocyte activation are ill defined. We show herein that LAT, once internalized, transits through the Golgi-trans-Golgi network (TGN), where it is repolarized to the immune synapse. This retrograde transport of LAT depends on the small GTPase Rab6 and the target soluble -ethylmaleimide-sensitive factor attachment protein receptor (t-SNARE) Syntaxin-16, two regulators of the endosome-to-Golgi/TGN retrograde transport. We also show in vitro in Syntaxin-16- or Rab6-silenced human cells and in vivo in CD4 T lymphocytes of the Rab6 knockout mouse that this retrograde traffic controls TCR stimulation. These results establish that the retrograde traffic of LAT from the plasma membrane to the Golgi-TGN controls the polarized delivery of LAT at the immune synapse and T lymphocyte activation.

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Année de publication : 2017

Anna Sawicka, Avin Babataheri, Stéphanie Dogniaux, Abdul I Barakat, David Gonzalez-Rodriguez, Claire Hivroz, Julien Husson (2017 Sep 22)

Micropipette Force Probe to quantify single-cell force generation: application to T cell activation.

Molecular biology of the cell : DOI : mbc.E17-06-0385 En savoir plus
Résumé

In response to engagement of surface molecules, cells generate active forces that regulate many cellular processes. Developing tools that permit gathering mechanical and morphological information on these forces is of the utmost importance. Here we describe a new technique, the Micropipette Force Probe, that uses a micropipette as a flexible cantilever that can aspirate at its tip a bead that is coated with molecules of interest and is brought in contact with the cell. This technique simultaneously allows tracking the resulting changes in cell morphology and mechanics as well as measuring the forces generated by the cell. To illustrate the power of this technique, we applied it to the study of human primary T lymphocytes (T cells). It allowed the fine monitoring of pushing and pulling forces generated by T cells in response to various activating antibodies and bending stiffness of the micropipette. We further dissected the sequence of mechanical and morphological events occurring during T cell activation to model force generation and to reveal heterogeneity in the cell population studied. We also report the first measurement of the changes in Young’s modulus of T cells during their activation, showing that T cells stiffen within the first minutes of the activation process.

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Michael Saitakis, Stéphanie Dogniaux, Christel Goudot, Nathalie Bufi, Sophie Asnacios, Mathieu Maurin, Clotilde Randriamampita, Atef Asnacios, Claire Hivroz (2017 Jun 9)

Different TCR-induced T lymphocyte responses are potentiated by stiffness with variable sensitivity.

eLife : DOI : 10.7554/eLife.23190 En savoir plus
Résumé

T cells are mechanosensitive but the effect of stiffness on their functions is still debated. We characterize herein how human primary CD4(+) T cell functions are affected by stiffness within the physiological Young’s modulus range of 0.5 kPa to 100 kPa. Stiffness modulates T lymphocyte migration and morphological changes induced by TCR/CD3 triggering. Stiffness also increases TCR-induced immune system, metabolism and cell-cycle-related genes. Yet, upon TCR/CD3 stimulation, while cytokine production increases within a wide range of stiffness, from hundreds of Pa to hundreds of kPa, T cell metabolic properties and cell cycle progression are only increased by the highest stiffness tested (100 kPa). Finally, mechanical properties of adherent antigen-presenting cells modulate cytokine production by T cells. Together, these results reveal that T cells discriminate between the wide range of stiffness values found in the body and adapt their responses accordingly.

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Claire Hivroz, Paola Larghi, Mabel Jouve, Laurence Ardouin (2017 Mar 4)

Purification of LAT-Containing Membranes from Resting and Activated T Lymphocytes.

Methods in molecular biology (Clifton, N.J.) : 355-368 : DOI : 10.1007/978-1-4939-6881-7_21 En savoir plus
Résumé

In T lymphocytes, the immune synapse is an active zone of vesicular traffic. Directional transport of vesicular receptors and signaling molecules from or to the immune synapse has been shown to play an important role in T-cell receptor (TCR) signal transduction. However, how vesicular trafficking is regulating the activation of T cells is still a burning question, and the characterization of these intracellular compartments remains the first step to understand this process. We describe herein a protocol, which combines a separation of membranes on flotation gradient with an affinity purification of Strep-tagged fusion transmembrane proteins with Strep-Tactin(®) resin, allowing the purification of membranes containing the Strep-tagged molecule of interest. By keeping the membranes intact, this protocol leads to the purification of molecules physically associated with the Strep-tagged protein as well as of molecules present in the same membrane compartment: transmembrane proteins, proteins strongly associated with the membranes, and luminal proteins. The example shown herein is the purification of membrane compartment prepared from T lymphocytes expressing LAT fused to a Strep-tag.

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Année de publication : 2016

Asma Beldi-Ferchiou, Marion Lambert, Stéphanie Dogniaux, Frédéric Vély, Eric Vivier, Daniel Olive, Stéphanie Dupuy, Frank Levasseur, David Zucman, Céleste Lebbé, Damien Sène, Claire Hivroz, Sophie Caillat-Zucman (2016 Sep 24)

PD-1 mediates functional exhaustion of activated NK cells in patients with Kaposi sarcoma.

Oncotarget : DOI : 10.18632/oncotarget.12150 En savoir plus
Résumé

Programmed Death-1 (PD-1), an inhibitory receptor expressed by activated lymphocytes, is involved in regulating T- and B-cell responses. PD-1 and its ligands are exploited by a variety of cancers to facilitate tumor escape through PD-1-mediated functional exhaustion of effector T cells. Here, we report that PD-1 is upregulated on Natural Killer (NK) cells from patients with Kaposi sarcoma (KS). PD-1 was expressed in a sub-population of activated, mature CD56dimCD16pos NK cells with otherwise normal expression of NK surface receptors. PD-1pos NK cells from KS patients were hyporesponsive ex vivo following direct triggering of NKp30, NKp46 or CD16 activating receptors, or short stimulation with NK cell targets. PD-1pos NK cells failed to degranulate and release IFNγ, but exogenous IL-2 or IL-15 restored this defect. That PD-1 contributed to NK cell functional impairment and was not simply a marker of dysfunctional NK cells was confirmed in PD-1-transduced NKL cells. In vitro, PD-1 was induced at the surface of healthy control NK cells upon prolonged contact with cells expressing activating ligands, i.e. a condition mimicking persistent stimulation by tumor cells. Thus, PD-1 appears to plays a critical role in mediating NK cell exhaustion. The existence of this negative checkpoint fine-tuning NK activation highlights the possibility that manipulation of the PD-1 pathway may be a strategy for circumventing tumor escape not only from the T cell-, but also the NK-cell mediated immune surveillance.

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Lionel Guillou, Avin Babataheri, Michael Saitakis, Armelle Bohineust, Stéphanie Dogniaux, Claire Hivroz, Abdul I Barakat, Julien Husson (2016 Sep 9)

T lymphocyte passive deformation is controlled by unfolding of membrane surface reservoirs.

Molecular biology of the cell : DOI : mbc.E16-06-0414 En savoir plus
Résumé

T lymphocytes in the human body routinely undergo large deformations, both passively when going through narrow capillaries and actively when transmigrating across endothelial cells or squeezing through tissue. We investigate physical factors that enable and limit such deformations and explore how passive and active deformations may differ. Employing micropipette aspiration to mimic squeezing through narrow capillaries, we find that T lymphocytes maintain a constant volume while increasing their apparent membrane surface area upon aspiration. Human resting T lymphocytes, T lymphoblasts and the leukemic Jurkat T cells all exhibit membrane rupture above a critical membrane area expansion that is independent of either micropipette size or aspiration pressure. The unfolded membrane matches the excess membrane contained in microvilli and membrane folds, as determined using scanning electron microscopy. In contrast, during transendothelial migration, a form of active deformation, we find that the membrane surface exceeds by a factor of two the amount of membrane stored in microvilli and folds. These results suggest that internal membrane reservoirs need to be recruited, possibly through exocytosis, for large active deformations to occur.

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Chantal Lagresle-Peyrou, Sonia Luce, Farid Ouchani, Tayebeh Shabi Soheili, Hanem Sadek, Myriam Chouteau, Amandine Durand, Isabelle Pic, Jacek Majewski, Chantal Brouzes, Nathalie Lambert, Armelle Bohineust, Els Verhoeyen, François-Loïc Cosset, Aude Magerus-Chatinet, Frédéric Rieux-Laucat, Virginie Gandemer, Delphine Monnier, Catherine Heijmans, Marielle van Gijn, Virgil A Dalm, Nizar Mahlaoui, Jean-Louis Stephan, Capucine Picard, Anne Durandy, Sven Kracker, Claire Hivroz, Nada Jabado, Geneviève de Saint Basile, Alain Fischer, Marina Cavazzana, Isabelle André-Schmutz (2016 Jul 14)

X-linked primary immunodeficiency associated with hemizygous mutations in the moesin (MSN) gene.

The Journal of allergy and clinical immunology : DOI : S0091-6749(16)30423-7 En savoir plus
Résumé

We investigated 7 male patients (from 5 different families) presenting with profound lymphopenia, hypogammaglobulinemia, fluctuating monocytopenia and neutropenia, a poor immune response to vaccine antigens, and increased susceptibility to bacterial and varicella zoster virus infections.

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Roshni Basu, Benjamin M Whitlock, Julien Husson, Audrey Le Floc'h, Weiyang Jin, Alon Oyler-Yaniv, Farokh Dotiwala, Gregory Giannone, Claire Hivroz, Nicolas Biais, Judy Lieberman, Lance C Kam, Morgan Huse (2016 Mar 1)

Cytotoxic T Cells Use Mechanical Force to Potentiate Target Cell Killing.

Cell : 100-10 : DOI : 10.1016/j.cell.2016.01.021 En savoir plus
Résumé

The immunological synapse formed between a cytotoxic T lymphocyte (CTL) and an infected or transformed target cell is a physically active structure capable of exerting mechanical force. Here, we investigated whether synaptic forces promote the destruction of target cells. CTLs kill by secreting toxic proteases and the pore forming protein perforin into the synapse. Biophysical experiments revealed a striking correlation between the magnitude of force exertion across the synapse and the speed of perforin pore formation on the target cell, implying that force potentiates cytotoxicity by enhancing perforin activity. Consistent with this interpretation, we found that increasing target cell tension augmented pore formation by perforin and killing by CTLs. Our data also indicate that CTLs coordinate perforin release and force exertion in space and time. These results reveal an unappreciated physical dimension to lymphocyte function and demonstrate that cells use mechanical forces to control the activity of outgoing chemical signals.

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Claire Hivroz, Michael Saitakis (2016 Feb 26)

Biophysical Aspects of T Lymphocyte Activation at the Immune Synapse.

Frontiers in immunology : 46 : DOI : 10.3389/fimmu.2016.00046 En savoir plus
Résumé

T lymphocyte activation is a pivotal step of the adaptive immune response. It requires the recognition by T-cell receptors (TCR) of peptides presented in the context of major histocompatibility complex molecules (pMHC) present at the surface of antigen-presenting cells (APCs). T lymphocyte activation also involves engagement of costimulatory receptors and adhesion molecules recognizing ligands on the APC. Integration of these different signals requires the formation of a specialized dynamic structure: the immune synapse. While the biochemical and molecular aspects of this cell-cell communication have been extensively studied, its mechanical features have only recently been addressed. Yet, the immune synapse is also the place of exchange of mechanical signals. Receptors engaged on the T lymphocyte surface are submitted to many tensile and traction forces. These forces are generated by various phenomena: membrane undulation/protrusion/retraction, cell mobility or spreading, and dynamic remodeling of the actomyosin cytoskeleton inside the T lymphocyte. Moreover, the TCR can both induce force development, following triggering, and sense and convert forces into biochemical signals, as a bona fide mechanotransducer. Other costimulatory molecules, such as LFA-1, engaged during immune synapse formation, also display these features. Moreover, T lymphocytes themselves are mechanosensitive, since substrate stiffness can modulate their response. In this review, we will summarize recent studies from a biophysical perspective to explain how mechanical cues can affect T lymphocyte activation. We will particularly discuss how forces are generated during immune synapse formation; how these forces affect various aspects of T lymphocyte biology; and what are the key features of T lymphocyte response to stiffness.

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Année de publication : 2015

Omar I Vivar, Giulia Masi, Jean-Marie Carpier, Joao G Magalhaes, Donatella Galgano, Gregory J Pazour, Sebastian Amigorena, Claire Hivroz, Cosima T Baldari (2015 Dec 31)

IFT20 controls LAT recruitment to the immune synapse and T-cell activation in vivo.

Proceedings of the National Academy of Sciences of the United States of America : 386-91 : DOI : 10.1073/pnas.1513601113 En savoir plus
Résumé

Biogenesis of the immune synapse at the interface between antigen-presenting cells and T cells assembles and organizes a large number of membrane proteins required for effective signaling through the T-cell receptor. We showed previously that the intraflagellar transport protein 20 (IFT20), a component of the intraflagellar transport system, controls polarized traffic during immune synapse assembly. To investigate the role of IFT20 in primary CD4(+) T cells in vitro and in vivo, we generated mice bearing a conditional defect of IFT20 expression in T cells. We show that in the absence of IFT20, although cell spreading and the polarization of the centrosome were unaffected, T-cell receptor (TCR)-mediated signaling and recruitment of the signaling adaptor LAT (linker for activation of T cells) at the immune synapse were reduced. As a consequence, CD4(+) T-cell activation and proliferation were also defective. In vivo, conditional IFT20-deficient mice failed to mount effective antigen-specific T-cell responses, and their T cells failed to induce colitis after adoptive transfer to Rag(-/-) mice. IFT20 is therefore required for the delivery of the intracellular pool of LAT to the immune synapse in naive primary T lymphocytes and for effective T-cell responses in vivo.

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