ERM-Dependent Assembly of T Cell Receptor Signaling and Co-stimulatory Molecules on Microvilli prior to Activation

Ghosh, Shirsendu; Bartolo, Vincenzo Di; Tubul, Liron; Shimoni, Eyal; Kartvelishvily, Elena; Dadosh, Tali; Feigelson, Sara W.; Alon, Ronen; Alcover, Andrés; Haran, Gilad

doi:10.1016/j.celrep.2020.02.069

Cited by 60 publications

(93 citation statements)

References 100 publications

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“…Recent studies showed that CD45 segregates from isolated tight microvilli contacts within seconds of TCR engagement, and that TCR phosphorylation negatively correlates with this TCR-CD45 separation (Razvag et al, 2018;Razvag et al, 2019). Moreover, a recent study showed that the TCR coreceptors, CD4 and CD2, and early signaling molecules, Lck and LAT, are highly enriched on microvilli (Ghosh et al, 2020). Together, these findings implicate microvilli as specialized cellular domains where initial antigen recognition and signal transduction occur.…”

Section: Discussionmentioning

confidence: 97%

CD45 pre-exclusion from the tips of microvilli establishes a phosphatase-free zone for early TCR triggering

Jung

Wen

Ley

2020

Preprint

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Graphical abstract 2 SummaryThe tyrosine phosphatase CD45 is a major gatekeeper for restraining T cell activation. Its exclusion from the immunological synapse (IS) is crucial for TCR signal transduction. Here, we used expansion superresolution microscopy to reveal that CD45 is pre-excluded from the tips of microvilli on primary T cells prior to antigen encounter. This pre-exclusion was diminished by depleting cholesterol or by engineering the transmembrane domain of CD45 to increase its membrane integration length, but was independent of the CD45 extracellular domain. We further show that brief microvilli-mediated contacts can induce Ca 2+ influx in mouse antigen-specific T cells engaged by antigen-pulsed APCs. We propose that the absence of CD45 phosphatase activity at the tips of microvilli enables or facilitates TCR triggering from brief T cell-APC contacts before formation of a stable IS, and that these microvilli-mediated contacts represent the earliest step in the initiation of a T cell adaptive immune response.

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Section: Discussionmentioning

confidence: 97%

CD45 pre-exclusion from the tips of microvilli establishes a phosphatase-free zone for early TCR triggering

Jung

Wen

Ley

2020

Preprint

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“…We further showed that TCR localization on microvilli is mediated by ERM (ezrin, radixin and moesin) proteins, which connect membrane proteins to the cortical actin cytoskeleton. Our study has established the role of microvilli as specialized activation hubs of T cells 9 . Interestingly, recent work suggests that microvillar protrusions do not disappear following T-cell activation and do not collapse during maturation of the immunological synapse (IS).…”

Section: Introductionmentioning

confidence: 76%

“…A different role for microvilli has been recently suggested by observations that lymphocytes use their microvilli to search for antigenic signals presented by rare antigen presenting cells (APCs) 3,9,10 . Moreover, microvilli have been shown to be involved in force-driven penetration of the dense glycocalyx barrier of APCs, which may otherwise hinder ligand/receptor interaction.…”

Section: Introductionmentioning

confidence: 99%

“…While our experiments were conducted on fixed cells, experiments by Krummel and coworkers 3 using light-sheet microscopy and by Sherman and coworkers 12 using a combination of atomic force and super-resolution microscopies demonstrated the dynamic nature of microvillar involvement in contact formation with APCs. Recently, we used the MC approach to show that, in addition to TCRs, multiple membrane proteins involved in the initiation of the immune response are also enriched on microvilli 9 . We further showed that TCR localization on microvilli is mediated by ERM (ezrin, radixin and moesin) proteins, which connect membrane proteins to the cortical actin cytoskeleton.…”

Section: Introductionmentioning

confidence: 99%

“…A careful comparison to pairs of proteins that are not known to interact facilitates this kind of analysis. A detailed description of our CP analysis is, however, beyond the scope of this Protocol, and can be found in our recent publication 9 .…”

Section: Introductionmentioning

confidence: 99%

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Microvillar cartography: a super-resolution single-molecule imaging method to map the positions of membrane proteins with respect to cellular surface topography

Ghosh

Alon

Alcover

et al. 2020

Preprint

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We introduce Microvillar Cartography (MC), a method to map proteins on cellular surfaces with respect to the membrane topography. The surfaces of many cells are not smooth, but are rather covered with various protrusions such as microvilli. These protrusions may play key roles in multiple cellular functions, due to their ability to control the distribution of specific protein assemblies on the cell surface. Thus, for example, we have shown that the T-cell receptor and several of its proximal signaling proteins reside on microvilli, while others are excluded from these projections. These results have indicated that microvilli can function as key signaling hubs for the initiation of the immune response. MC has facilitated our observations of particular surface proteins and their specialized distribution on microvillar and non-microvillar compartments. MC combines membrane topography imaging, using variable-angle total internal microscopy, with stochastic localization nanoscopy, which generates deep subdiffraction maps of protein distribution. Since the method is based on light microscopy, it avoids some of the pitfalls inherent to electron-microscopy-based techniques, such as dehydration, carbon coating and immunogold clustering, and is amenable to future developments involving e.g. live-cell imaging. This Protocol details the procedures we developed for MC, which can be readily adopted to study a broad range of cell surface molecules and dissect their distribution within distinct surface assemblies under multiple cell activation states.

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The T Cell Journey: A Tour de Force

et al. 2022

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T cells act as the puppeteers in the adaptive immune response, and their dysfunction leads to the initiation and progression of pathological conditions. During their lifetime, T cells experience myriad forces that modulate their effector functions. These forces are imposed by interacting cells, surrounding tissues, and shear forces from fluid movement. In this review, a journey with T cells is made, from their development to their unique characteristics, including the early studies that uncovered their mechanosensitivity. Then the studies pertaining to the responses of T cell activation to changes in antigen‐presenting cells’ physical properties, to their immediate surrounding extracellular matrix microenvironment, and flow conditions are highlighted. In addition, it is explored how pathological conditions like the tumor microenvironment can hinder T cells and allow cancer cells to escape elimination.

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ERM-Dependent Assembly of T Cell Receptor Signaling and Co-stimulatory Molecules on Microvilli prior to Activation

Cited by 60 publications

References 100 publications

CD45 pre-exclusion from the tips of microvilli establishes a phosphatase-free zone for early TCR triggering

CD45 pre-exclusion from the tips of microvilli establishes a phosphatase-free zone for early TCR triggering

Microvillar cartography: a super-resolution single-molecule imaging method to map the positions of membrane proteins with respect to cellular surface topography

The T Cell Journey: A Tour de Force

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