Interfacial actin protrusions mechanically enhance killing by cytotoxic T cells

Tamzalit, Fella; Wang, Mitchell S.; Jin, Weiyang; Tello-Lafoz, Maria; Boyko, Vitaly; Heddleston, John M.; Black, Charles T.; Kam, Lance C.; Huse, Morgan

doi:10.1126/sciimmunol.aav5445

Interfacial actin protrusions mechanically enhance killing by cytotoxic T cells

Journal Article · Fri Mar 22 04:00:00 EDT 2019 · Science Immunology

DOI:https://doi.org/10.1126/sciimmunol.aav5445· OSTI ID:1508745

^[1]; ^[2]; ^[3]; ^[1]; Boyko, Vitaly ^[1]; ^[4]; ^[5]; ^[3]; ^[1]

Memorial Sloan Kettering Cancer Center, New York, NY (United States)
Weill Cornell Medical College, New York, NY (United States)
Columbia Univ., New York, NY (United States)
Howard Hughes Medical Institute, Ashburn, VA (United States)
Brookhaven National Lab. (BNL), Upton, NY (United States)

Cytotoxic T lymphocytes (CTLs) kill by forming immunological synapses with target cells and secreting toxic proteases and the pore-forming protein perforin into the intercellular space. Immunological synapses are highly dynamic structures that boost perforin activity by applying mechanical force against the target cell. Here, we used high-resolution imaging and microfabrication to investigate how CTLs exert synaptic forces and coordinate their mechanical output with perforin secretion. Using micropatterned stimulatory substrates that enable synapse growth in three dimensions, we found that perforin release occurs at the base of actin-rich protrusions that extend from central and intermediate locations within the synapse. These protrusions, which depended on the cytoskeletal regulator WASP and the Arp2/3 actin nucleation complex, were required for synaptic force exertion and efficient killing. They also mediated physical deformation of the target cell surface during CTL–target cell interactions. Here our results reveal the mechanical basis of cellular cytotoxicity and highlight the functional importance of dynamic, three-dimensional architecture in immune cell-cell interfaces.

View Accepted Manuscript (DOE)

Research Organization:: Brookhaven National Laboratory (BNL), Upton, NY (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)

Grant/Contract Number:: SC0012704

OSTI ID:: 1508745

Report Number(s):: BNL--211580-2019-JAAM

Journal Information:: Science Immunology, Journal Name: Science Immunology Journal Issue: 33 Vol. 4; ISSN 2470-9468

Publisher:: AAASCopyright Statement

Country of Publication:: United States

Language:: English

References (64)

Wiskott-Aldrich syndrome protein: Emerging mechanisms in immunity Rivers, Elizabeth; Thrasher, Adrian J. European Journal of Immunology, Vol. 47, Issue 11 https://doi.org/10.1002/eji.201646715	journal	September 2017
Dynamic Actin Polymerization Drives T Cell Receptor–Induced Spreading Bunnell, Stephen C.; Kapoor, Veena; Trible, Ronald P. Immunity, Vol. 14, Issue 3 https://doi.org/10.1016/S1074-7613(01)00112-1	journal	March 2001
The Immunological Synapse of CTL Contains a Secretory Domain and Membrane Bridges Stinchcombe, Jane C.; Bossi, Giovanna; Booth, Sarah Immunity, Vol. 15, Issue 5 https://doi.org/10.1016/S1074-7613(01)00234-5	journal	November 2001
Wiskott-Aldrich Syndrome Protein Regulates Lipid Raft Dynamics during Immunological Synapse Formation Dupré, Loı̈c; Aiuti, Alessandro; Trifari, Sara Immunity, Vol. 17, Issue 2 https://doi.org/10.1016/S1074-7613(02)00360-6	journal	August 2002
Visualizing Dynamic Microvillar Search and Stabilization during Ligand Detection by T Cells Cai, En; Marchuk, Kyle; Beemiller, Peter Biophysical Journal, Vol. 114, Issue 3 https://doi.org/10.1016/j.bpj.2017.11.2992	journal	February 2018
Opposing Effects of PKCθ and WASp on Symmetry Breaking and Relocation of the Immunological Synapse Sims, Tasha N.; Soos, Timothy J.; Xenias, Harry S. Cell, Vol. 129, Issue 4 https://doi.org/10.1016/j.cell.2007.03.037	journal	May 2007
Cytotoxic T Cells Use Mechanical Force to Potentiate Target Cell Killing Basu, Roshni; Whitlock, Benjamin M.; Husson, Julien Cell, Vol. 165, Issue 1 https://doi.org/10.1016/j.cell.2016.01.021	journal	March 2016
The Wiskott-Aldrich Syndrome Protein Contributes to the Assembly of the LFA-1 Nanocluster Belt at the Lytic Synapse Houmadi, Raïssa; Guipouy, Delphine; Rey-Barroso, Javier Cell Reports, Vol. 22, Issue 4 https://doi.org/10.1016/j.celrep.2017.12.088	journal	January 2018
The WAVE2 Complex Regulates Actin Cytoskeletal Reorganization and CRAC-Mediated Calcium Entry during T Cell Activation Nolz, Jeffrey C.; Gomez, Timothy S.; Zhu, Peimin Current Biology, Vol. 16, Issue 1 https://doi.org/10.1016/j.cub.2005.11.036	journal	January 2006
Role for the Abi/Wave Protein Complex in T Cell Receptor-Mediated Proliferation and Cytoskeletal Remodeling Zipfel, Patricia A.; Bunnell, Stephen C.; Witherow, D. Scott Current Biology, Vol. 16, Issue 1 https://doi.org/10.1016/j.cub.2005.12.024	journal	January 2006
Distinct Roles for the Actin Nucleators Arp2/3 and hDia1 during NK-Mediated Cytotoxicity Butler, Boyd; Cooper, John A. Current Biology, Vol. 19, Issue 22 https://doi.org/10.1016/j.cub.2009.10.029	journal	December 2009
Distinct Roles for the Actin Nucleators Arp2/3 and hDia1 during NK-Mediated Cytotoxicity Butler, Boyd; Cooper, John A. Current Biology, Vol. 20, Issue 18 https://doi.org/10.1016/j.cub.2010.09.002	journal	September 2010
Formins Regulate the Actin-Related Protein 2/3 Complex-Independent Polarization of the Centrosome to the Immunological Synapse Gomez, Timothy S.; Kumar, Karan; Medeiros, Ricardo B. Immunity, Vol. 26, Issue 2 https://doi.org/10.1016/j.immuni.2007.01.008	journal	February 2007
Transcellular Diapedesis Is Initiated by Invasive Podosomes Carman, Christopher V.; Sage, Peter T.; Sciuto, Tracey E. Immunity, Vol. 26, Issue 6 https://doi.org/10.1016/j.immuni.2007.04.015	journal	June 2007
T Cell Costimulation via the Integrin VLA-4 Inhibits the Actin-Dependent Centralization of Signaling Microclusters Containing the Adaptor SLP-76 Nguyen, Ken; Sylvain, Nicholas R.; Bunnell, Stephen C. Immunity, Vol. 28, Issue 6 https://doi.org/10.1016/j.immuni.2008.04.019	journal	June 2008
Kinetics of Early T Cell Receptor Signaling Regulate the Pathway of Lytic Granule Delivery to the Secretory Domain Beal, Allison M.; Anikeeva, Nadia; Varma, Rajat Immunity, Vol. 31, Issue 4 https://doi.org/10.1016/j.immuni.2009.09.004	journal	October 2009
Actin Depletion Initiates Events Leading to Granule Secretion at the Immunological Synapse Ritter, Alex T.; Asano, Yukako; Stinchcombe, Jane C. Immunity, Vol. 42, Issue 5 https://doi.org/10.1016/j.immuni.2015.04.013	journal	May 2015
Cytotoxic T lymphocyte mediated lysis without release of serine esterase Ostergaard, Hanne L.; Kane, Kevin P.; Mescher, Matthew F. Nature, Vol. 330, Issue 6143 https://doi.org/10.1038/330071a0	journal	November 1987
Centrosome polarization delivers secretory granules to the immunological synapse Stinchcombe, Jane C.; Majorovits, Endre; Bossi, Giovanna Nature, Vol. 443, Issue 7110 https://doi.org/10.1038/nature05071	journal	September 2006
Characterization of two classes of small molecule inhibitors of Arp2/3 complex Nolen, B. J.; Tomasevic, N.; Russell, A. Nature, Vol. 460, Issue 7258 https://doi.org/10.1038/nature08231	journal	August 2009
T cell killing does not require the formation of a stable mature immunological synapse Purbhoo, Marco A.; Irvine, Darrell J.; Huppa, Johannes B. Nature Immunology, Vol. 5, Issue 5 https://doi.org/10.1038/ni1058	journal	March 2004
Imaging techniques for assaying lymphocyte activation in action Balagopalan, Lakshmi; Sherman, Eilon; Barr, Valarie A. Nature Reviews Immunology, Vol. 11, Issue 1 https://doi.org/10.1038/nri2903	journal	December 2010
The ARP2/3 complex: an actin nucleator comes of age Goley, Erin D.; Welch, Matthew D. Nature Reviews Molecular Cell Biology, Vol. 7, Issue 10 https://doi.org/10.1038/nrm2026	journal	October 2006
Cytotoxic T lymphocytes kill multiple targets simultaneously via spatiotemporal uncoupling of lytic and stimulatory synapses Wiedemann, A.; Depoil, D.; Faroudi, M. Proceedings of the National Academy of Sciences, Vol. 103, Issue 29 https://doi.org/10.1073/pnas.0600651103	journal	July 2006
Mechanisms for segregating T cell receptor and adhesion molecules during immunological synapse formation in Jurkat T cells Kaizuka, Y.; Douglass, A. D.; Varma, R. Proceedings of the National Academy of Sciences, Vol. 104, Issue 51 https://doi.org/10.1073/pnas.0710258105	journal	December 2007
CD4+ T-cell synapses involve multiple distinct stages Ueda, H.; Morphew, M. K.; McIntosh, J. R. Proceedings of the National Academy of Sciences, Vol. 108, Issue 41 https://doi.org/10.1073/pnas.1113703108	journal	September 2011
Diacylglycerol promotes centrosome polarization in T cells via reciprocal localization of dynein and myosin II Liu, X.; Kapoor, T. M.; Chen, J. K. Proceedings of the National Academy of Sciences, Vol. 110, Issue 29 https://doi.org/10.1073/pnas.1306180110	journal	July 2013
CD28 and CD3 have complementary roles in T-cell traction forces Bashour, Keenan T.; Gondarenko, Alexander; Chen, Haoqian Proceedings of the National Academy of Sciences, Vol. 111, Issue 6 https://doi.org/10.1073/pnas.1315606111	journal	January 2014
Three-dimensional localization of T-cell receptors in relation to microvilli using a combination of superresolution microscopies Jung, Yunmin; Riven, Inbal; Feigelson, Sara W. Proceedings of the National Academy of Sciences, Vol. 113, Issue 40 https://doi.org/10.1073/pnas.1605399113	journal	September 2016
Cortical actin recovery at the immunological synapse leads to termination of lytic granule secretion in cytotoxic T lymphocytes Ritter, Alex T.; Kapnick, Senta M.; Murugesan, Sricharan Proceedings of the National Academy of Sciences, Vol. 114, Issue 32 https://doi.org/10.1073/pnas.1710751114	journal	July 2017
The WAVE2 complex regulates T cell receptor signaling to integrins via Abl- and CrkL–C3G-mediated activation of Rap1 Nolz, Jeffrey C.; Nacusi, Lucas P.; Segovis, Colin M. The Journal of Cell Biology, Vol. 182, Issue 6 https://doi.org/10.1083/jcb.200801121	journal	September 2008
Formin-generated actomyosin arcs propel T cell receptor microcluster movement at the immune synapse Murugesan, Sricharan; Hong, Jinsung; Yi, Jason The Journal of Cell Biology, Vol. 215, Issue 3 https://doi.org/10.1083/jcb.201603080	journal	October 2016
Annular PIP ₃ accumulation controls actin architecture and modulates cytotoxicity at the immunological synapse Le Floc'h, Audrey; Tanaka, Yoshihiko; Bantilan, Niels S. The Journal of Cell Biology, Vol. 203, Issue 3 https://doi.org/10.1083/jcb.2033oia133	journal	November 2013
Antigen Receptor–Induced Activation and Cytoskeletal Rearrangement Are Impaired in Wiskott-Aldrich Syndrome Protein–Deficient Lymphocytes Zhang, Jinyi; Shehabeldin, Amro; da Cruz, Luis A. G. The Journal of Experimental Medicine, Vol. 190, Issue 9 https://doi.org/10.1084/jem.190.9.1329	journal	November 1999
Annular PIP ₃ accumulation controls actin architecture and modulates cytotoxicity at the immunological synapse Le Floc’h, Audrey; Tanaka, Yoshihiko; Bantilan, Niels S. The Journal of Experimental Medicine, Vol. 210, Issue 12 https://doi.org/10.1084/jem.20131324	journal	November 2013
The WAVE2 complex regulates T cell receptor signaling to integrins via Abl- and CrkL–C3G-mediated activation of Rap1 Nolz, Jeffrey C.; Nacusi, Lucas P.; Segovis, Colin M. The Journal of Experimental Medicine, Vol. 205, Issue 10 https://doi.org/10.1084/jem20510oia21	journal	September 2008
Annular PIP ₃ accumulation controls actin architecture and modulates cytotoxicity at the immunological synapse Le Floc'h, Audrey; Tanaka, Yoshihiko; Bantilan, Niels S. The Journal of General Physiology, Vol. 142, Issue 6 https://doi.org/10.1085/jgp.1426oia47	journal	November 2013
Actin retrograde flow and actomyosin II arc contraction drive receptor cluster dynamics at the immunological synapse in Jurkat T cells Yi, Jason; Wu, Xufeng S.; Crites, Travis Molecular Biology of the Cell, Vol. 23, Issue 5 https://doi.org/10.1091/mbc.E11-08-0731	journal	March 2012
Actin retrograde flow and actomyosin II arc contraction drive receptor cluster dynamics at the immunological synapse in Jurkat T cells Yi, Jason; Wu, Xufeng S.; Crites, Travis Molecular Biology of the Cell, Vol. 23, Issue 5 https://doi.org/10.1091/mbc.e11-08-0731	journal	March 2012
Cell:cell interactions in the immune system Batista, Facundo D.; Dustin, Michael L. Immunological Reviews, Vol. 251, Issue 1 https://doi.org/10.1111/imr.12025	journal	December 2012
Cytotoxic immunological synapses: NK and CTL synapses Dustin, Michael L.; Long, Eric O. Immunological Reviews, Vol. 235, Issue 1 https://doi.org/10.1111/j.0105-2896.2010.00904.x	journal	April 2010
Wiskott-Aldrich syndrome: a comprehensive review: Wiskott-Aldrich syndrome Massaad, Michel J.; Ramesh, Narayanaswamy; Geha, Raif S. Annals of the New York Academy of Sciences, Vol. 1285, Issue 1 https://doi.org/10.1111/nyas.12049	journal	March 2013
Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution Chen, Bi-Chang; Legant, Wesley R.; Wang, Kai Science, Vol. 346, Issue 6208 https://doi.org/10.1126/science.1257998	journal	October 2014
Visualizing dynamic microvillar search and stabilization during ligand detection by T cells Cai, En; Marchuk, Kyle; Beemiller, Peter Science, Vol. 356, Issue 6338 https://doi.org/10.1126/science.aal3118	journal	May 2017
Diacylglycerol kinase establishes T cell polarity by shaping diacylglycerol accumulation at the immunological synapse Chauveau, A.; Le Floc'h, A.; Bantilan, N. S. Science Signaling, Vol. 7, Issue 340 https://doi.org/10.1126/scisignal.2005287	journal	August 2014
Secretory Mechanisms in Cell-Mediated Cytotoxicity Stinchcombe, Jane C.; Griffiths, Gillian M. Annual Review of Cell and Developmental Biology, Vol. 23, Issue 1 https://doi.org/10.1146/annurev.cellbio.23.090506.123521	journal	November 2007
How Do Cytotoxic Lymphocytes Kill Cancer Cells? Martinez-Lostao, L.; Anel, A.; Pardo, J. Clinical Cancer Research, Vol. 21, Issue 22 https://doi.org/10.1158/1078-0432.CCR-15-0685	journal	November 2015
Cytoskeletal remodeling mediated by WASp in dendritic cells is necessary for normal immune synapse formation and T-cell priming Bouma, Gerben; Mendoza-Naranjo, Ariadna; Blundell, Michael P. Blood, Vol. 118, Issue 9 https://doi.org/10.1182/blood-2011-03-340265	journal	September 2011
Abnormalities of follicular helper T-cell number and function in Wiskott-Aldrich syndrome Zhang, Xuan; Dai, Rongxin; Li, Wenyan Blood, Vol. 127, Issue 25 https://doi.org/10.1182/blood-2015-06-652636	journal	June 2016
Wiskott-Aldrich syndrome protein is necessary for efficient IgG-mediated phagocytosis Lorenzi, Roberto; Brickell, Paul M.; Katz, David R. Blood, Vol. 95, Issue 9 https://doi.org/10.1182/blood.V95.9.2943.009k17_2943_2946	journal	May 2000
The Wiskott-Aldrich syndrome protein regulates CTL cytotoxicity and is required for efficient killing of B cell lymphoma targets De Meester, Julie; Calvez, Ronan; Valitutti, Salvatore Journal of Leukocyte Biology, Vol. 88, Issue 5 https://doi.org/10.1189/jlb.0410197	journal	August 2010
WASp-dependent actin cytoskeleton stability at the dendritic cell immunological synapse is required for extensive, functional T cell contacts Malinova, Dessislava; Fritzsche, Marco; Nowosad, Carla R. Journal of Leukocyte Biology, Vol. 99, Issue 5 https://doi.org/10.1189/jlb.2A0215-050RR	journal	November 2015
Natural Killer Cell Lytic Granule Secretion Occurs through a Pervasive Actin Network at the Immune Synapse Rak, Gregory D.; Mace, Emily M.; Banerjee, Pinaki P. PLoS Biology, Vol. 9, Issue 9 https://doi.org/10.1371/journal.pbio.1001151	journal	September 2011
Remodelling of Cortical Actin Where Lytic Granules Dock at Natural Killer Cell Immune Synapses Revealed by Super-Resolution Microscopy Brown, Alice C. N.; Oddos, Stephane; Dobbie, Ian M. PLoS Biology, Vol. 9, Issue 9 https://doi.org/10.1371/journal.pbio.1001152	journal	September 2011
Force Generation upon T Cell Receptor Engagement Husson, Julien; Chemin, Karine; Bohineust, Armelle PLoS ONE, Vol. 6, Issue 5 https://doi.org/10.1371/journal.pone.0019680	journal	May 2011
The Variable Hinge Region of Novel PKCs Determines Localization to Distinct Regions of the Immunological Synapse Basu, Roshni; Chen, Yuedan; Quann, Emily J. PLoS ONE, Vol. 9, Issue 4 https://doi.org/10.1371/journal.pone.0095531	journal	April 2014
Actin retrograde flow and actomyosin II arc contraction drive receptor cluster dynamics at the immunological synapse in Jurkat T cells Travis, Crites,; Jason, Yi,; A., Hammer III, John The University of North Carolina at Chapel Hill University Libraries https://doi.org/10.17615/790z-gn50	text	January 2012
Cortical actin recovery at the immunological synapse leads to termination of lytic granule secretion in cytotoxic T lymphocytes. Ritter, Alex T.; Kapnick, Senta M.; Murugesan, Sricharan Apollo - University of Cambridge Repository https://doi.org/10.17863/cam.30628	text	January 2017
The Wiskott-Aldrich syndrome protein permits assembly of a focused immunological synapse enabling sustained T-cell receptor signaling Calvez, R.; Lafouresse, F.; De Meester, J. Haematologica, Vol. 96, Issue 10 https://doi.org/10.3324/haematol.2011.040204	journal	June 2011
Docking of Lytic Granules at the Immunological Synapse in Human CTL Requires Vti1b-Dependent Pairing with CD3 Endosomes Qu, Bin; Pattu, Varsha; Junker, Christian The Journal of Immunology, Vol. 186, Issue 12 https://doi.org/10.4049/jimmunol.1003471	journal	May 2011
Antigen Recognition Is Facilitated by Invadosome-like Protrusions Formed by Memory/Effector T Cells Sage, Peter T.; Varghese, Laya M.; Martinelli, Roberta The Journal of Immunology, Vol. 188, Issue 8 https://doi.org/10.4049/jimmunol.1102594	journal	March 2012
Cutting Edge: The Wiskott-Aldrich Syndrome Protein Is Required for Efficient Phagocytosis of Apoptotic Cells Leverrier, Yann; Lorenzi, Roberto; Blundell, Michael P. The Journal of Immunology, Vol. 166, Issue 8 https://doi.org/10.4049/jimmunol.166.8.4831	journal	April 2001
Differential Roles for Wiskott-Aldrich Syndrome Protein in Immune Synapse Formation and IL-2 Production Cannon, Judy L.; Burkhardt, Janis K. The Journal of Immunology, Vol. 173, Issue 3 https://doi.org/10.4049/jimmunol.173.3.1658	journal	July 2004
Actin foci facilitate activation of the phospholipase C-γ in primary T lymphocytes via the WASP pathway Kumari, Sudha; Depoil, David; Martinelli, Roberta eLife, Vol. 4 https://doi.org/10.7554/eLife.04953	journal	March 2015

Cited By (4)

Loss of ARPC1B impairs cytotoxic T lymphocyte maintenance and cytolytic activity. Randzavola, Lyra O.; Strege, Katharina; Juzans, Marie Apollo - University of Cambridge Repository https://doi.org/10.17863/cam.46931	text	January 2019
Actin Cytoskeleton Straddling the Immunological Synapse between Cytotoxic Lymphocytes and Cancer Cells Wurzer, Hannah; Hoffmann, Céline; Al Absi, Antoun Cells, Vol. 8, Issue 5 https://doi.org/10.3390/cells8050463	journal	May 2019
Mechanotransduction in T Cell Development, Differentiation and Function Rushdi, Muaz; Li, Kaitao; Yuan, Zhou Cells, Vol. 9, Issue 2 https://doi.org/10.3390/cells9020364	journal	February 2020
Loss of ARPC1B impairs cytotoxic T lymphocyte maintenance and cytolytic activity Randzavola, Lyra O.; Strege, Katharina; Juzans, Marie Journal of Clinical Investigation, Vol. 129, Issue 12 https://doi.org/10.1172/jci129388	journal	October 2019

Similar Records

Coronin 3 involvement in F-actin-dependent processes at the cell cortex

Journal Article · Fri Mar 09 23:00:00 EST 2007 · Experimental Cell Research · OSTI ID:20972126

Cell confinement reveals a branched-actin independent circuit for neutrophil polarity

Journal Article · Thu Oct 10 00:00:00 EDT 2019 · PLoS Biology (Online) · OSTI ID:1599812

Cytotoxic T lymphocyte-dependent tumor growth inhibition by a vascular endothelial growth factor-superantigen conjugate

Journal Article · Fri Nov 02 00:00:00 EDT 2012 · Biochemical and Biophysical Research Communications · OSTI ID:22210315

Related Subjects

36 MATERIALS SCIENCE

Interfacial actin protrusions mechanically enhance killing by cytotoxic T cells

Citation Formats

References (64)

Cited By (4)

Similar Records

Related Subjects