Durable Interactions of T Cells With T Cell Receptor Stimuli in the Absence of a Stable Immunological Synapse

Mayya, Viveka; Judokusumo, Edward; Neiswanger, Willie; Depoil, David; Blair, David A.; Wiggins, Chris H.; Kam, Lance C.; Dustin, Michael L.

doi:10.2139/ssrn.3155633

Cited by 6 publications

(8 citation statements)

References 28 publications

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“…The difference in behavior may imply that autoreactive Th17 cells in the CNS have a higher Ca 2+ threshold for stopping than naïve antigen-specific T cells in secondary lymphoid organs. Our observations further raise the possibility that Th17 cell reactivation in situ may occur through kinapses in which T cells remain motile and exhibit Ca 2+ signals while contacting APCs (55,62,63).…”

Section: Discussionmentioning

confidence: 59%

Regulatory T cells suppress Th17 cell Ca ²⁺ signaling in the spinal cord during murine autoimmune neuroinflammation

Othy

Jairaman

Dynes

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

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T lymphocyte motility and interaction dynamics with other immune cells are vital determinants of immune responses. Regulatory T (Treg) cells prevent autoimmune disorders by suppressing excessive lymphocyte activity, but how interstitial motility patterns of Treg cells limit neuroinflammation is not well understood. We used two-photon microscopy to elucidate the spatial organization, motility characteristics, and interactions of endogenous Treg and Th17 cells together with antigen-presenting cells (APCs) within the spinal cord leptomeninges in experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis. Th17 cells arrive before the onset of clinical symptoms, distribute uniformly during the peak, and decline in numbers during later stages of EAE. In contrast, Treg cells arrive after Th17 cells and persist during the chronic phase. Th17 cells meander widely, interact with APCs, and exhibit cytosolic Ca2+ transients and elevated basal Ca2+ levels before the arrival of Treg cells. In contrast, Treg cells adopt a confined, repetitive-scanning motility while contacting APCs. These locally confined but highly motile Treg cells limit Th17 cells from accessing APCs and suppress Th17 cell Ca2+ signaling by a mechanism that is upstream of store-operated Ca2+ entry. Finally, Treg cell depletion increases APC numbers in the spinal cord and exaggerates ongoing neuroinflammation. Our results point to fundamental differences in motility characteristics between Th17 and Treg cells in the inflamed spinal cord and reveal three potential cellular mechanisms by which Treg cells regulate Th17 cell effector functions: reduction of APC density, limiting access of Th17 cells to APCs, and suppression of Th17 Ca2+ signaling.

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Section: Discussionmentioning

confidence: 59%

Regulatory T cells suppress Th17 cell Ca ²⁺ signaling in the spinal cord during murine autoimmune neuroinflammation

Othy

Jairaman

Dynes

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

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“…Within seconds, T cells seeded onto an APS spread to form a symmetric contact interface (Appendix Fig S1). Symmetric synapses persisted for~10 min, followed by a transition period when the cells began polarizing and becoming motile (Negulescu et al, 1996;Hons et al, 2018;Houmadi et al, 2018;Mayya et al, 2018), reflected in an increase in AR (Fig 1B,Movies EV1 and EV2). Structured illumination microscopy (SIM) revealed the presence of peripheral actin-rich lamellipodia and punctate actin structures distributed across the synaptic interface ( Fig 1C), termed "actin foci" (Kumari et al, 2015) in stable (5 min) synapse.…”

Section: Resultsmentioning

confidence: 99%

“…To model synapse formation and eventual disengagement, we seeded mouse primary naïve CD4 + primary T cells (referred to as “T cells” henceforth) onto an anti‐CD3/intercellular adhesion molecule‐1 (ICAM‐1)‐coated coverslip (antigen‐presenting surface, APS) and allowed synapses to form. We then assessed cellular polarization over time by recording the cells’ morphologic aspect ratio, AR, because the contact interface elongation, reflected as increase in AR, is tightly linked to the motile state of the T cell (Hons et al , ; Houmadi et al , ; Mayya et al , ; Negulescu et al , ; Fig A, and Appendix Figs S1 and S2). That is, synapses are radially symmetric in their stable state and polarize to break symmetry thereby acquiring an elongated morphology prior to resumption of motility (Movies EV1 and EV2).…”

Section: Resultsmentioning

confidence: 99%

Cytoskeletal tension actively sustains the migratory T‐cell synaptic contact

et al. 2020

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When migratory T cells encounter antigen‐presenting cells (APCs), they arrest and form radially symmetric, stable intercellular junctions termed immunological synapses which facilitate exchange of crucial biochemical information and are critical for T‐cell immunity. While the cellular processes underlying synapse formation have been well characterized, those that maintain the symmetry, and thereby the stability of the synapse, remain unknown. Here we identify an antigen‐triggered mechanism that actively promotes T‐cell synapse symmetry by generating cytoskeletal tension in the plane of the synapse through focal nucleation of actin via Wiskott–Aldrich syndrome protein (WASP), and contraction of the resultant actin filaments by myosin II. Following T‐cell activation, WASP is degraded, leading to cytoskeletal unraveling and tension decay, which result in synapse breaking. Thus, our study identifies and characterizes a mechanical program within otherwise highly motile T cells that sustains the symmetry and stability of the T cell–APC synaptic contact.

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“…Interestingly, these images show that the T cell mechanically scanned a significant fraction of antigen (81 ± 28% over its initial contact area, n = 10 cells) within a 10-min migration time window. Thus, the dynamic T cell synapse (kinapse) represents a zone of TCR mechanosensing (19)(20)(21). Upon addition of the unlocking strand, probes reset back to the real-time state and exclusively showed tension at the trailing cell edge (unlocked state; Fig.…”

Section: Resultsmentioning

confidence: 99%

DNA probes that store mechanical information reveal transient piconewton forces applied by T cells

Kellner

Pui‐Yan

et al. 2019

Proc. Natl. Acad. Sci. U.S.A.

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The advent of molecular tension probes for real-time mapping of piconewton forces in living systems has had a major impact on mechanobiology. For example, DNA-based tension probes have revealed roles for mechanics in platelet, B cell, T cell, and fibroblast function. Nonetheless, imaging short-lived forces transmitted by low-abundance receptors remains a challenge. This is a particular problem for mechanoimmunology where ligand–receptor bindings are short lived, and a few antigens are sufficient for cell triggering. Herein, we present a mechanoselection strategy that uses locking oligonucleotides to preferentially and irreversibly bind DNA probes that are mechanically strained over probes at rest. Thus, infrequent and short-lived mechanical events are tagged. This strategy allows for integration and storage of mechanical information into a map of molecular tension history. Upon addition of unlocking oligonucleotides that drive toehold-mediated strand displacement, the probes reset to the real-time state, thereby erasing stored mechanical information. As a proof of concept, we applied this strategy to study OT-1 T cells, revealing that the T cell receptor (TCR) mechanically samples antigens carrying single amino acid mutations. Such events are not detectable using conventional tension probes. Each mutant peptide ligand displayed a different level of mechanical sampling and spatial scanning by the TCR that strongly correlated with its functional potency. Finally, we show evidence that T cells transmit pN forces through the programmed cell death receptor-1 (PD1), a major target in cancer immunotherapy. We anticipate that mechanical information storage will be broadly useful in studying the mechanobiology of the immune system.

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Durable Interactions of T Cells With T Cell Receptor Stimuli in the Absence of a Stable Immunological Synapse

Cited by 6 publications

References 28 publications

Regulatory T cells suppress Th17 cell Ca ²⁺ signaling in the spinal cord during murine autoimmune neuroinflammation

Regulatory T cells suppress Th17 cell Ca ²⁺ signaling in the spinal cord during murine autoimmune neuroinflammation

Cytoskeletal tension actively sustains the migratory T‐cell synaptic contact

DNA probes that store mechanical information reveal transient piconewton forces applied by T cells

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Durable Interactions of T Cells With T Cell Receptor Stimuli in the Absence of a Stable Immunological Synapse

Cited by 6 publications

References 28 publications

Regulatory T cells suppress Th17 cell Ca 2+ signaling in the spinal cord during murine autoimmune neuroinflammation

Regulatory T cells suppress Th17 cell Ca 2+ signaling in the spinal cord during murine autoimmune neuroinflammation

Cytoskeletal tension actively sustains the migratory T‐cell synaptic contact

DNA probes that store mechanical information reveal transient piconewton forces applied by T cells

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Regulatory T cells suppress Th17 cell Ca ²⁺ signaling in the spinal cord during murine autoimmune neuroinflammation

Regulatory T cells suppress Th17 cell Ca ²⁺ signaling in the spinal cord during murine autoimmune neuroinflammation