Alexander A. Smoligovets's research while affiliated with University of California, Berkeley and other places
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Publications (13)
The T-cell actin cytoskeleton mediates adaptive immune system responses to peptide antigens by physically directing the motion and clustering of T-cell receptors (TCRs) on the cell surface. When TCR movement is impeded by externally applied physical barriers, the actin network exhibits transient enrichment near the trapped receptors. The coordinate...
The actin cytoskeleton provides a dynamic framework to support membrane organization and cellular signaling events. The importance of actin in T cell function has long been recognized to go well beyond the maintenance of cell morphology and transport of proteins. Over the past several years, our understanding of actin in T cell activation has expan...
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TCR translocation is coupled to actin retrograde flow. (A) Simultaneous total internal reflection fluorescence (TIRF) images of TCR labeled with αTCR Fab (Alexa Fluor 594) and EGFP-UtrCH during the formation of an immunological synapse in a control cell. (B) Time-averaged radial velocities, <V(t)>, of all TCR microclusters and EGFP-UtrCH in control...
TIRF images of TCR and pZAP-70 (pY 319) in T cells pretreated with DMSO, blebbistatin, or ML-7. T cells were fixed at the time indicated after the initial cell-bilayer contact.
(TIF)
Morphological quantification of the immunological synapse. TIRF images of TCRs labeled with H57 αTCR Fab (Alexa Fluor 594) and ICAM-1 (Alexa Fluor 488) are shown. Cells were pretreated with DMSO, blebbistatin, or ML-7, and fixed at (A) 3 min and (B) 10 min after interacting with bilayers. Normalized intensities of TCRs and ICAM-1 are plotted versus...
Activation of T cell receptor (TCR) by antigens occurs in concert with an elaborate multi-scale spatial reorganization of proteins at the immunological synapse, the junction between a T cell and an antigen-presenting cell (APC). The directed movement of molecules, which intrinsically requires physical forces, is known to modulate biochemical signal...
Physical constraints on TCR microcluster translocation impede actin retrograde flow. (A) TIRF, reflection interference contrast microscopy (RICM), and bright field (BF) images of T cells expressing EGFP-UtrCH on unpatterned or patterned bilayers. Scale bars: 5 µm. (B) Time-averaged radial velocities (<V(t)>) of EGFP- UtrCH in individual cells are p...
Colocalization of TCR and pCasL in T cells pretreated with DMSO, blebbistatin, or ML-7. The percentages of TCR microclusters colocalized with pCasL are shown for the indicated stimulation times prior to fixation. Each column is an averaged value from approximately 200 cells. Data were reproduced in 2 independent experiments.
(TIF)
T cell triggering through T-cell antigen receptors (TCRs) results in spatial assembly of the receptors on multiple length scales. This assembly is mediated by the T cell actin cytoskeleton, which reorganizes in response to TCR phosphorylation and then induces the coalescence of TCRs into microclusters, followed by their unification into a micromete...
Activation of T cell receptor (TCR) by antigens occurs in concert with an elaborate multi-scale spatial reorganization of proteins at the immunological synapse, the junction between a T cell and an antigen-presenting cell (APC). Signaling through discrete T cell receptors (TCRs) in the context of immunological synapse, involves the orchestrated mov...
Photoactivatable fluorescent proteins offer the possibility to optically tag and track the location of molecules in their bright state with high spatial and temporal resolution. Several reports of patterned photoactivation have emerged since the development of a photoactivatable variant of the green fluorescent protein (PaGFP) and the demonstration...
Citations
... TCRs specifically recognize rare agonist peptide-major histocompatibility complexes (pMHCs) among numerous self-pMHCs on the surface of the cell being surveyed to trigger adaptive immune responses and therefore are essential molecules to combat infection and cancer (Chakraborty and Weiss, 2014;van der Merwe and Dushek, 2011). TCRs are known to form microclusters on the surface of the T cell and represent a key mechanism toward understanding T-cell signaling and function (Campi et al., 2005;Crites et al., 2014;Gagnon et al., 2012;Hashimoto-Tane et al., 2016, 2011Hu et al., 2016;Huang et al., 2013;Hui et al., 2017;Lewis et al., 2018;Lillemeier et al., 2010;Murugesan et al., 2016;Roh et al., 2015;Sasmal et al., 2020;Schamel et al., 2005;Smoligovets et al., 2012;Taylor et al., 2017;Varma et al., 2006;Wang et al., 2019b;Yi et al., 2019;Yokosuka et al., 2005). ...
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... Low-defect, fluid supported lipid bilayers (SLB) were prepared from a solution of small unilamellar vesicles (SUVs). SUVs were first prepared by mixing 95% DOPC (1,2-dioleoyl-sn- T cell harvesting and culture CD4+ T cells expressing the AND TCR (51) were harvested, cultured, and transduced as previously described (15,52). Briefly, T cells were harvested from hemizygous transgenic mice from the cross of (B10.Cg-Tg(TcrAND)53Hed/J) x (B10.BR-H2k2 H2-T18a/SgSnJ) strains (The Jackson Laboratory, Bar Harbor, ME) and activated by moth cytochrome c peptide (MCC 88-103 ) immediately after harvest. ...
... This decays as CD4 + T cell Wiskott-Aldrich syndrome protein (WASP) is degraded, consistent with previous findings on the role of WASP in APC-T cell IS stability (56). Effector cell membrane tension may be monitored by the co-localized mechanosensory protein CasL, which is phosphorylated on conformational stretching (75,94). This represents effector cell programmed release of the CD4 T cell stimulatory synapse. ...
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... The use of nanopatterned supported lipid bilayer substrates with physical barriers uncovered a requirement for nucleation and micron-scale assembly of E-cadherin for a-catenin activation (22). Nanopatterned substrates have been successfully employed to elucidate the role of receptor clustering in a number of cellular signaling systems, including T-cell activation, EphA2 receptor tyrosine kinase signaling, and integrin adhesion (24)(25)(26)(60)(61)(62)(63)(64)(65)(66). Unlike genetic or pharmacological perturbations that may have offtarget or deleterious effects (e.g., on protein stability), these nanopatterned substrates allow receptor clustering to be perturbed in a purely physical way, thereby avoiding such effects. ...
... These clusters merge at the center of the IS zone and are referred to as the central supramolecular cluster. In contrast, microclusters found at the periphery of the synapse join to form a highly contractile zone called the peripheral supramolecular activation cluster (271). ...
