TCR Down-Regulation Controls T Cell Homeostasis

Boding, Lasse; Bonefeld, Charlotte M.; Nielsen, Bodil; Lauritsen, Jens Peter H.; Essen, Marina Rode von; Hansen, Anker; Larsen, Jeppe Madura; Nielsen, Morten Milek; Ødum, Niels; Geisler, Carsten

doi:10.4049/jimmunol.0901539

Cited by 13 publications

(13 citation statements)

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“…This phenomenon is of particular relevance when redirecting T cells with defined specificity towards target cells. While reduced response of early-stage CD45RO + T cells is due to down-regulated expression of TCR components, especially the CD3-gamma chain, following ligand triggering [19], we here revealed that T cells in more differentiated stages, in particular in the KLRG-1 −/low CD57 − CD7 + intermediate and KLRG-1 + CD57 + CD7 − late-stage of terminal differentiation, express the TCR and bind specific tetramers with similar efficiencies; CD7 − T cells, however, respond less efficiently with IFN-gamma and cytolytic degranulation than the CD7 + KLRG-1 −/low CD57 − T cells. While hypo-responsiveness of intermediate-stage T cells is due to blockade through PD-1/PD-1L interaction, this is not the case for late-stage T cells (unpublished data).…”

Section: Discussionmentioning

confidence: 99%

The CD3-Zeta Chimeric Antigen Receptor Overcomes TCR Hypo-Responsiveness of Human Terminal Late-Stage T Cells

et al. 2012

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Adoptive therapy of malignant diseases with tumor-specific cytotoxic T cells showed remarkable efficacy in recent trials. Repetitive T cell receptor (TCR) engagement of target antigen, however, inevitably ends up in hypo-responsive cells with terminally differentiated KLRG-1+ CD57+ CD7− phenotype limiting their therapeutic efficacy. We here revealed that hypo-responsiveness of CMV-specific late-stage CD8+ T cells is due to reduced TCR synapse formation compared to younger cells. Membrane anchoring of TCR components contributes to T cell hypo-responsiveness since dislocation of galectin-3 from the synapse by swainsonine restored both TCR synapse formation and T cell response. Transgenic expression of a CD3-zeta signaling chimeric antigen receptor (CAR) recovered hypo-responsive T cells to full effector functions indicating that the defect is restricted to TCR membrane components while synapse formation of the transgenic CAR was not blocked. CAR engineered late-stage T cells released cytokines and mediated redirected cytotoxicity as efficiently as younger effector T cells. Our data provide a rationale for TCR independent, CAR mediated activation in the adoptive cell therapy to avoid hypo-responsiveness of late-stage T cells upon repetitive antigen encounter.

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Section: Discussionmentioning

confidence: 99%

The CD3-Zeta Chimeric Antigen Receptor Overcomes TCR Hypo-Responsiveness of Human Terminal Late-Stage T Cells

et al. 2012

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“…For murine FACS analyses of cell surface and intracellular molecules, we used antibodies against CD8 (53-6.7), TCRVβ8 (F23.1), TCRVα2 (B20.1), CD62L (MEL-14), CD44 (IM7), IFN-γ (XMG1.2), and TNF-α (MP6.XT22) all from BD Biosciences, granzyme B (GB11, Invitrogen) and perforin (eBioOMAK-D, eBioscience). For phenotyping, cells from thymus, spleen, and LNs were isolated, stained with antibodies against CD4 (RM4-5), CD8 (53-6.7), TCRVα2 (B20.1), and CD19 (1D3) from BD Biosciences, and analyzed by FACS as previously described [44]. For CD107a staining CTLs were restimulated for the time indicated with 100 ng/mL gp [33][34][35][36][37][38][39][40][41] in the presence of CD107a-FITC antibody (1D4B from BD Biosciences) and 3 μM monensin as previously described [29].…”

Section: Expansion Of T Cells and Facs Analysismentioning

confidence: 99%

Midline 1 directs lytic granule exocytosis and cytotoxicity of mouse killer T cells

et al. 2014

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Midline 1 (MID1) is a microtubule-associated ubiquitin ligase that regulates protein phosphatase 2A activity. Loss-of-function mutations in MID1 lead to the X-linked Opitz G/BBB syndrome characterized by defective midline development during embryogenesis. Here, we show that MID1 is strongly upregulated in murine cytotoxic lymphocytes (CTLs), and that it controls TCR signaling, centrosome trafficking, and exocytosis of lytic granules. In accordance, we find that the killing capacity of MID1 −/− CTLs is impaired. Transfection of MID1 into MID1 −/− CTLs completely rescued lytic granule exocytosis, and vice versa, knockdown of MID1 inhibited exocytosis of lytic granules in WT CTLs, cementing a central role for MID1 in the regulation of granule exocytosis. Thus, MID1 orchestrates multiple events in CTL responses, adding a novel level of regulation to CTL activation and cytotoxicity.Keywords: Cytotoxicity r Granule exocytosis r Midline 1 r T cellsAdditional supporting information may be found in the online version of this article at the publisher's web-site IntroductionCytotoxic lymphocytes (CTLs) play a key role in the defense against infections. The CTL recognizes specific antigen/MHC complexes on infected target cells by the TCR. Specific recognition of a target cell leads to establishment of a stable conjugation, the immunological synapse (IS), between the CTL and the target cell [1][2][3]. Immediately after recognition of the target cell, the CTL polarizes its secretory machinery toward the IS. The centrosome, which in CTLs is often called the Correspondence: Dr. Carsten Geisler e-mail: cge@sund.ku.dk microtubule-organizing center (MTOC), relocates to the IS [4,5], and lytic granules that contain perforin and granzymes move along the microtubules toward the MTOC [6]. Here, they dock at the plasma membrane and deliver their content by exocytosis into the small secretory cleft of the IS [3,7,8]. Perforin and granzymes subsequently act in concert to induce apoptosis of the target cell [9]. Thus, the microtubule cytoskeleton plays a key role in directional secretion of lytic granules and thereby in CTL-mediated target cell killing [8]. It is still not clear how CTLs regulate reorganization of the microtubule cytoskeleton and exocytosis of lytic granules during encounter with target cells.C 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.eji-journal.eu 3110 Lasse Boding et al. Eur. J. Immunol. 2014. 44: 3109-3118 Midline 1 (MID1) is a ubiquitin ligase that belongs to the RBCC/TRIM family, which contains a RING domain followed by B-boxes and a coiled-coil domain [10,11]. MID1 is associated with the microtubules [12,13] and regulates the level of microtubuleassociated protein phosphatase 2A (PP2A) [14,15]. The X-linked Opitz G/BBB syndrome (OS) is caused by mutations of MID1 and is characterized by disorders that primarily affect the ventral midline [16]. Manifestations include ocular hypertelorism, genitourinary defects, clefts of palate and lip, trachea-esophageal fistulas, imperforate anus, and mental retard...

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“…Thus, during the steady state, TCR molecules transit through a recycling pool, as a result of internalization, endocytosis and cycling from and to the plasma membrane, via vesicular transport (Clark and Griffiths, 2003; Das et al, 2004; Liu et al, 2000; Minami et al, 1987; Myers et al, 2005). It has been speculated that constant TCR recycling during the steady-state, together with other biochemical events, contribute to control T cell homeostasis (Boding et al, 2009). In addition, the rapid cycling of surface receptors allows T cells to respond rapidly to stimuli by controlling responsiveness to peptide/MHC ligands (Harding and Unanue, 1990).…”

Section: Discussionmentioning

confidence: 99%

Inducible turnover of optineurin regulates T cell activation

Montecalvo

Watkins

Kane

2017

Molecular Immunology

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Optineurin (Optn) is an adaptor protein with homology to NF-κB essential-modulator (NEMO), the regulatory subunit of the IκB kinase (IKK) complex. Dysregulation of Optn has been linked to neurodegenerative, autoimmune and bone diseases. Optn shares a high degree of homology with NEMO, but is not part of the same high-molecular weight complex containing IKKα and IKKβ. Despite its homology with NEMO and the fact that it has been the subject of extensive study in several cell types, there are no published studies addressing the role of Optn during T cell activation. Here we demonstrate that ectopic expression of Optn down-regulates TCR- induced NF-κB activation and TNF-α production, in a manner dependent on ubiquitin-binding. Conversely, knock-down of Optn enhances NF-κB activation and the production of TNF-α. Consistent with a negative regulatory role for this protein, we observed transient loss of Optn after TCR stimulation in both cell lines and in primary murine T cells. The acute loss of Optn appears to be due to both protein degradation and exocytosis, the latter via activation-induced exosomes. This study therefore provides novel information regarding the role of Optn during TCR activation, suggesting the possible importance of Optn during inflammation and/or autoimmune diseases.

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TCR Down-Regulation Controls T Cell Homeostasis

Abstract: Material

Cited by 13 publications

References 62 publications

The CD3-Zeta Chimeric Antigen Receptor Overcomes TCR Hypo-Responsiveness of Human Terminal Late-Stage T Cells

The CD3-Zeta Chimeric Antigen Receptor Overcomes TCR Hypo-Responsiveness of Human Terminal Late-Stage T Cells

Midline 1 directs lytic granule exocytosis and cytotoxicity of mouse killer T cells

Inducible turnover of optineurin regulates T cell activation

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