scholarly journals Phosphoproteomics of CD2 signaling reveals AMPK-dependent regulation of lytic granule polarization in cytotoxic T cells

2020 ◽  
Vol 13 (631) ◽  
pp. eaaz1965 ◽  
Author(s):  
Vanessa Zurli ◽  
Tommaso Montecchi ◽  
Raphael Heilig ◽  
Isabel Poschke ◽  
Michael Volkmar ◽  
...  
Keyword(s):  
T Cells ◽  
Cytotoxic T Cells ◽  
Critical Role ◽  
Underlying Mechanism ◽  
Cell Receptor ◽  
Microtubule Organizing Center ◽  
Lytic Granules ◽  
Organizing Center ◽  
Lytic Granule ◽  
Insight Into

Understanding the costimulatory signaling that enhances the activity of cytotoxic T cells (CTLs) could identify potential targets for immunotherapy. Here, we report that CD2 costimulation plays a critical role in target cell killing by freshly isolated human CD8+ T cells, which represent a challenging but valuable model to gain insight into CTL biology. We found that CD2 stimulation critically enhanced signaling by the T cell receptor in the formation of functional immune synapses by promoting the polarization of lytic granules toward the microtubule-organizing center (MTOC). To gain insight into the underlying mechanism, we explored the CD2 signaling network by phosphoproteomics, which revealed 616 CD2-regulated phosphorylation events in 373 proteins implicated in the regulation of vesicular trafficking, cytoskeletal organization, autophagy, and metabolism. Signaling by the master metabolic regulator AMP-activated protein kinase (AMPK) was a critical node in the CD2 network, which promoted granule polarization toward the MTOC in CD8+ T cells. Granule trafficking was driven by active AMPK enriched on adjacent lysosomes, revealing previously uncharacterized signaling cross-talk between vesicular compartments in CD8+ T cells. Our results thus establish CD2 signaling as key for mediating cytotoxic killing and granule polarization in freshly isolated CD8+ T cells and strengthen the rationale to choose CD2 and AMPK as therapeutic targets to enhance CTL activity.

scholarly journals Phosphoproteomics of CD2 signaling reveals an AMPK-dependent regulation of lytic granule polarization in cytotoxic T cells

2019 ◽  
Author(s):  
Vanessa Zurli ◽  
Tommaso Montecchi ◽  
Raphael Heilig ◽  
Isabel Poschke ◽  
Michael Volkmar ◽  
...  
Keyword(s):  
T Cells ◽  
Cytotoxic T Cells ◽  
Critical Role ◽  
Cell Receptor ◽  
Microtubule Organizing Center ◽  
Lytic Granules ◽  
Cytoskeleton Organization ◽  
Depth Analysis ◽  
Organizing Center ◽  
Insight Into

SummaryThe in-depth analysis of costimulatory signaling enhancing the activity of cytotoxic T cells (CTLs) represents a major approach towards immunotherapy development. Here we report that CD2 costimulation plays a critical role in killing by freshly isolated human CTLs, which represent a challenging but valuable study model to gain insight into CTL biology. We show that CD2 triggering critically aids signaling by the T cell receptor in the formation of functional immune synapses by promoting the polarization of lytic granules towards the microtubule-organizing center (MTOC). To gain insight into the underlying elusive mechanism, we explored the CD2 signaling network by phosphoproteomics, which revealed 616 CD2-regulated phosphorylation events in 373 proteins implicated in the regulation of vesicular trafficking, cytoskeleton organization, autophagy and metabolism. Strikingly, signaling by the master metabolic regulator AMP-activated protein kinase (AMPK) represents a functionally critical node of the CD2 network which regulates granule polarization towards the MTOC in CTLs. Granule trafficking is driven by active AMPK enriched on adjacent lysosomes, illustrating a novel signaling cross-talk between vesicular compartments in CTLs. Our results thus establish CD2 signaling as key for regulating cytotoxic killing and granule polarization in freshly isolated CTLs and strengthens the rationale to choose CD2 and AMPK as therapeutic targets to boost CTL activity.

LOSS OF CYTOTOXIC T LYMPHOCYTE FUNCTION IN CHEDIAK-HIGASHI SYNDROME ARISES FROM A SECRETORY DEFECT THAT PREVENTS LYTIC GRANULE EXOCYTOSIS

PEDIATRICS ◽  
1996 ◽  
Vol 98 (2) ◽  
pp. 349-349
Author(s):  
Kathleen E. Sullivan
Keyword(s):  
T Cells ◽  
T Lymphocyte ◽  
Cytotoxic T Lymphocyte ◽  
Cytotoxic T Cells ◽  
Molecular Defect ◽  
Lymphocyte Function ◽  
Lytic Granules ◽  
Hematopoietic Lineage ◽  
Chediak Higashi Syndrome ◽  
Lytic Granule

Cytotoxic T cells from a patient with chediak-Higashi syndrome formed giant lytic granules that were not secreted in response to stimulation. This led to abnormal cytolytic function. The authors conclude that the molecular defect in Chediak-Higashi syndrome lies in a protein that is generally involved in vesicle fusion and is crucial for the secretion of lysosomal contents for cells of the hematopoietic lineage.

scholarly journals Engineering CD4+ T Cells to Enhance Cancer Immunity

Cells ◽  
2020 ◽  
Vol 9 (7) ◽  
pp. 1721 ◽  
Author(s):  
Francesca Sillito ◽  
Angelika Holler ◽  
Hans J. Stauss
Keyword(s):  
T Cells ◽  
T Cell ◽  
Mhc Class Ii ◽  
Cd4 T Cells ◽  
Cytotoxic T Cells ◽  
Critical Role ◽  
Cell Receptor ◽  
Convincing Evidence ◽  
Indirect Pathway ◽  
Tumour Immunity

This review presents key advances in combining T cell receptor (TCR) gene transfer to redirect T-cell specificity with gene engineering in order to enhance cancer-protective immune function. We discuss how emerging insights might be applied to CD4+ T cells. Although much attention has been paid to the role of CD8+ cytotoxic T cells in tumour protection, we provide convincing evidence that CD4+ helper T cells play a critical role in cancer immune responses in animal models and also in patients. We demonstrate that genetic engineering technologies provide exciting opportunities to extend the specificity range of CD4+ T cells from MHC class-II-presented epitopes to include peptides presented by MHC class I molecules. Functional enhancement of tumour immunity can improve the sensitivity of T cells to cancer antigens, promote survival in a hostile tumour microenvironment, boost cancer-protective effector mechanisms and enable the formation of T-cell memory. Engineered cancer-specific CD4+ T cells may contribute to protective immunity by a direct pathway involving cancer cell killing, and by an indirect pathway that boosts the function, persistence and memory formation of CD8+ T cells.

scholarly journals SRPS associated protein WDR60 regulates the multipolar-to-bipolar transition of migrating neurons during cortical development

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Cui Li ◽  
Yu Zheng ◽  
Yufang Zheng ◽  
Zhiheng Xu
Keyword(s):  
Critical Role ◽  
Underlying Mechanism ◽  
Stable Form ◽  
Microtubule Organizing Center ◽  
Microtubule Organization ◽  
Repeat Domain ◽  
Organizing Center ◽  
Newborn Neurons ◽  
And Migration ◽  
Cellular Components

AbstractMutations of WD40 repeat domain 60 (WDR60) have been identified in short-rib polydactyly syndromes (SRPS I–V), a group of lethal congenital disorders characterized by short ribs, polydactyly, and a range of extraskeletal phenotypes. However, the underlying mechanism is still unclear. Here, we report that WDR60 is essential for embryonic development and plays a critical role in the multipolar-bipolar transition and migration of newborn neurons during brain development. Mechanically, we found that WDR60 was located at the microtubule-organizing center to control microtubule organization and possibly, the trafficking of cellular components. Importantly, the migration defect caused by Wdr60 knockdown could be rescued by the stable form of α-Tubulin, α-TubulinK40Q (an acetylation-mimicking mutant). These findings identified a non-cilia function of WDR60 and provided insight into its biological function, as well as the pathogenesis of WDR60 deficiency associated with SRPS.

scholarly journals Terminal transport of lytic granules to the immune synapse is mediated by the kinesin-1/Slp3/Rab27a complex

Blood ◽  
2012 ◽  
Vol 119 (17) ◽  
pp. 3879-3889 ◽  
Author(s):  
Mathieu Kurowska ◽  
Nicolas Goudin ◽  
Nadine T. Nehme ◽  
Magali Court ◽  
Jérôme Garin ◽  
...  
Keyword(s):  
T Lymphocytes ◽  
Cytotoxic T Lymphocytes ◽  
Target Cells ◽  
Microtubule Organizing Center ◽  
Immune Synapse ◽  
Lytic Granules ◽  
Polarized Secretion ◽  
Organizing Center ◽  
Dependent Transport ◽  
Lytic Granule

Abstract Cytotoxic T lymphocytes kill target cells via the polarized secretion of cytotoxic granules at the immune synapse. The lytic granules are initially recruited around the polarized microtubule-organizing center. In a dynein-dependent transport process, the granules move along microtubules toward the microtubule-organizing center in the minus-end direction. Here, we found that a kinesin-1–dependent process is required for terminal transport and secretion of polarized lytic granule to the immune synapse. We show that synaptotagmin-like protein 3 (Slp3) is an effector of Rab27a in cytotoxic T lymphocytes and interacts with kinesin-1 through the tetratricopeptide repeat of the kinesin-1 light chain. Inhibition of the Rab27a/Slp3/kinesin-1 transport complex impairs lytic granule secretion. Our data provide further molecular insights into the key functional and regulatory mechanisms underlying the terminal transport of cytotoxic granules and the latter's secretion at the immune synapse.

scholarly journals The Msd1–Wdr8–Pkl1 complex anchors microtubule minus ends to fission yeast spindle pole bodies

2015 ◽  
Vol 209 (4) ◽  
pp. 549-562 ◽  
Author(s):  
Masashi Yukawa ◽  
Chiho Ikebe ◽  
Takashi Toda
Keyword(s):  
Fission Yeast ◽  
Critical Role ◽  
Spindle Pole Body ◽  
Spindle Pole ◽  
Microtubule Organizing Center ◽  
Spatiotemporal Regulation ◽  
Spindle Pole Bodies ◽  
Organizing Center ◽  
Spindle Microtubules ◽  
Insight Into

The minus ends of spindle microtubules are anchored to a microtubule-organizing center. The conserved Msd1/SSX2IP proteins are localized to the spindle pole body (SPB) and the centrosome in fission yeast and humans, respectively, and play a critical role in microtubule anchoring. In this paper, we show that fission yeast Msd1 forms a ternary complex with another conserved protein, Wdr8, and the minus end–directed Pkl1/kinesin-14. Individual deletion mutants displayed the identical spindle-protrusion phenotypes. Msd1 and Wdr8 were delivered by Pkl1 to mitotic SPBs, where Pkl1 was tethered through Msd1–Wdr8. The spindle-anchoring defect imposed by msd1/wdr8/pkl1 deletions was suppressed by a mutation of the plus end–directed Cut7/kinesin-5, which was shown to be mutual. Intriguingly, Pkl1 motor activity was not required for its anchoring role once targeted to the SPB. Therefore, spindle anchoring through Msd1–Wdr8–Pkl1 is crucial for balancing the Cut7/kinesin-5–mediated outward force at the SPB. Our analysis provides mechanistic insight into the spatiotemporal regulation of two opposing kinesins to ensure mitotic spindle bipolarity.

scholarly journals CD28-B7 interactions allow the induction of CD8+ cytotoxic T lymphocytes in the absence of exogenous help.

1993 ◽  
Vol 177 (6) ◽  
pp. 1791-1796 ◽  
Author(s):  
F A Harding ◽  
J P Allison
Keyword(s):  
T Cells ◽  
T Cell ◽  
T Cell Activation ◽  
Cell Activation ◽  
Cytotoxic T Cells ◽  
Interleukin 2 ◽  
Cell Receptor ◽  
Histocompatibility Complex ◽  
Necessary And Sufficient ◽  
Additional Antigen

The activation requirements for the generation of CD8+ cytotoxic T cells (CTL) are poorly understood. Here we demonstrate that in the absence of exogenous help, a CD28-B7 interaction is necessary and sufficient for generation of class I major histocompatibility complex-specific CTL. Costimulation is required only during the inductive phase of the response, and not during the effector phase. Transfection of the CD28 counter receptor, B7, into nonstimulatory P815 cells confers the ability to elicit P815-specific CTL, and this response can be inhibited by anti-CD28 Fab or by the chimeric B7-binding protein CTLA4Ig. Anti-CD28 monoclonal antibody (mAb) can provide a costimulatory signal to CD8+ T cells when the costimulatory capacity of splenic stimulators is destroyed by chemical fixation. CD28-mediated signaling provokes the release of interleukin 2 (IL-2) from the CD8+ CTL precursors, as anti-CD28 mAb could be substituted for by the addition of IL-2, and an anti-IL-2 mAb can block the generation of anti-CD28-induced CTL. CD4+ cells are not involved in the costimulatory response in the systems examined. We conclude that CD8+ T cell activation requires two signals: an antigen-specific signal mediated by the T cell receptor, and an additional antigen nonspecific signal provided via a CD28-B7 interaction.

Sign in / Sign up

Export Citation Format

Share Document