Supramolecular Attack Particles Are Autonomous Killing Entities Released from Cytotoxic T Cells

Overview

Journal Science

Specialty Science

Date 2020 May 9

PMID 32381591

Citations 79

Authors

S Balint

S Muller

R Fischer

B M Kessler

M Harkiolaki

S Valitutti

M L Dustin

Affiliations

Soon will be listed here.

Abstract

Cytotoxic T lymphocytes (CTLs) kill infected and cancerous cells. We detected transfer of cytotoxic multiprotein complexes, called supramolecular attack particles (SMAPs), from CTLs to target cells. SMAPs were rapidly released from CTLs and were autonomously cytotoxic. Mass spectrometry, immunochemical analysis, and CRISPR editing identified a carboxyl-terminal fragment of thrombospondin-1 as an unexpected SMAP component that contributed to target killing. Direct stochastic optical reconstruction microscopy resolved a cytotoxic core surrounded by a thrombospondin-1 shell of ~120 nanometer diameter. Cryo-soft x-ray tomography analysis revealed that SMAPs had a carbon-dense shell and were stored in multicore granules. We propose that SMAPs are autonomous extracellular killing entities that deliver cytotoxic cargo targeted by the specificity of shell components.

Citing Articles

Activation-induced thrombospondin-4 works with thrombospondin-1 to build cytotoxic supramolecular attack particles.

Cassioli C, Capitani N, Staton C, Schirra C, Finetti F, Onnis A Proc Natl Acad Sci U S A. 2025; 122(6):e2413866122.

PMID: 39903110 PMC: 11831147. DOI: 10.1073/pnas.2413866122.

Stoichiometric constraints for detection of EV-borne biomarkers in blood.

Zarovni N, Mladenovic D, Brambilla D, Panico F, Chiari M J Extracell Vesicles. 2025; 14(2):e70034.

PMID: 39901737 PMC: 11791308. DOI: 10.1002/jev2.70034.

Harnessing extracellular vesicle-mediated crosstalk between T cells and cancer cells for therapeutic applications.

Budayr O, Miller B, Nguyen J J Control Release. 2024; 378:266-280.

PMID: 39657892 PMC: 11830559. DOI: 10.1016/j.jconrel.2024.12.011.

Extracellular vesicles and nanoparticles at a glance.

Jeppesen D, Zhang Q, Coffey R J Cell Sci. 2024; 137(23).

PMID: 39641198 PMC: 11658686. DOI: 10.1242/jcs.260201.

Formin-like 1β phosphorylation at S1086 is necessary for secretory polarized traffic of exosomes at the immune synapse in Jurkat T lymphocytes.

Ruiz-Navarro J, Fernandez-Hermira S, Sanz-Fernandez I, Barbeito P, Navarro-Zapata A, Perez-Martinez A Elife. 2024; 13.

PMID: 39479958 PMC: 11527432. DOI: 10.7554/eLife.96942.

References

Choudhuri K, Llodra J, Roth E, Tsai J, Gordo S, Wucherpfennig K . Polarized release of T-cell-receptor-enriched microvesicles at the immunological synapse. Nature. 2014; 507(7490):118-23. PMC: 3949170. DOI: 10.1038/nature12951. View

Metkar S, Wang B, Aguilar-Santelises M, Raja S, Uhlin-Hansen L, Podack E . Cytotoxic cell granule-mediated apoptosis: perforin delivers granzyme B-serglycin complexes into target cells without plasma membrane pore formation. Immunity. 2002; 16(3):417-28. DOI: 10.1016/s1074-7613(02)00286-8. View

Froelich C, Orth K, Turbov J, Seth P, Gottlieb R, Babior B . New paradigm for lymphocyte granule-mediated cytotoxicity. Target cells bind and internalize granzyme B, but an endosomolytic agent is necessary for cytosolic delivery and subsequent apoptosis. J Biol Chem. 1996; 271(46):29073-9. DOI: 10.1074/jbc.271.46.29073. View

Cocucci E, Meldolesi J . Ectosomes and exosomes: shedding the confusion between extracellular vesicles. Trends Cell Biol. 2015; 25(6):364-72. DOI: 10.1016/j.tcb.2015.01.004. View

Law R, Lukoyanova N, Voskoboinik I, Caradoc-Davies T, Baran K, Dunstone M . The structural basis for membrane binding and pore formation by lymphocyte perforin. Nature. 2010; 468(7322):447-51. DOI: 10.1038/nature09518. View

Peters P, Geuze H, van der Donk H, Slot J, Griffith J, Stam N . Molecules relevant for T cell-target cell interaction are present in cytolytic granules of human T lymphocytes. Eur J Immunol. 1989; 19(8):1469-75. DOI: 10.1002/eji.1830190819. View

Dustin M, Starr T, Varma R, Thomas V . Supported planar bilayers for study of the immunological synapse. Curr Protoc Immunol. 2008; Chapter 18:18.13.1-18.13.35. DOI: 10.1002/0471142735.im1813s76. View

Oldenborg P, Gresham H, Lindberg F . CD47-signal regulatory protein alpha (SIRPalpha) regulates Fcgamma and complement receptor-mediated phagocytosis. J Exp Med. 2001; 193(7):855-62. PMC: 2193364. DOI: 10.1084/jem.193.7.855. View

Schneider G . Cryo X-ray microscopy with high spatial resolution in amplitude and phase contrast. Ultramicroscopy. 1998; 75(2):85-104. DOI: 10.1016/s0304-3991(98)00054-0. View

10.

Harkiolaki M, Darrow M, Spink M, Kosior E, Dent K, Duke E . Cryo-soft X-ray tomography: using soft X-rays to explore the ultrastructure of whole cells. Emerg Top Life Sci. 2021; 2(1):81-92. PMC: 7289011. DOI: 10.1042/ETLS20170086. View

11.

Balint S, Vilanova I, Alvarez A, Lakadamyali M . Correlative live-cell and superresolution microscopy reveals cargo transport dynamics at microtubule intersections. Proc Natl Acad Sci U S A. 2013; 110(9):3375-80. PMC: 3587250. DOI: 10.1073/pnas.1219206110. View

12.

Stinchcombe J, Bossi G, Booth S, Griffiths G . The immunological synapse of CTL contains a secretory domain and membrane bridges. Immunity. 2001; 15(5):751-61. DOI: 10.1016/s1074-7613(01)00234-5. View

13.

Bates M, Dempsey G, Chen K, Zhuang X . Multicolor super-resolution fluorescence imaging via multi-parameter fluorophore detection. Chemphyschem. 2012; 13(1):99-107. PMC: 3353744. DOI: 10.1002/cphc.201100735. View

14.

Peters P, Borst J, Oorschot V, Fukuda M, Krahenbuhl O, Tschopp J . Cytotoxic T lymphocyte granules are secretory lysosomes, containing both perforin and granzymes. J Exp Med. 1991; 173(5):1099-109. PMC: 2118839. DOI: 10.1084/jem.173.5.1099. View

15.

Martina J, Wu X, Catalfamo M, Sakamoto T, Yi C, Hammer 3rd J . Imaging of lytic granule exocytosis in CD8+ cytotoxic T lymphocytes reveals a modified form of full fusion. Cell Immunol. 2011; 271(2):267-79. PMC: 3407469. DOI: 10.1016/j.cellimm.2011.07.004. View

16.

Misenheimer T, Mosher D . Calcium ion binding to thrombospondin 1. J Biol Chem. 1995; 270(4):1729-33. DOI: 10.1074/jbc.270.4.1729. View

17.

Masson D, Tschopp J . A family of serine esterases in lytic granules of cytolytic T lymphocytes. Cell. 1987; 49(5):679-85. DOI: 10.1016/0092-8674(87)90544-7. View

18.

Kalos M, Levine B, Porter D, Katz S, Grupp S, Bagg A . T cells with chimeric antigen receptors have potent antitumor effects and can establish memory in patients with advanced leukemia. Sci Transl Med. 2011; 3(95):95ra73. PMC: 3393096. DOI: 10.1126/scitranslmed.3002842. View

19.

Keefe D, Shi L, Feske S, Massol R, Navarro F, Kirchhausen T . Perforin triggers a plasma membrane-repair response that facilitates CTL induction of apoptosis. Immunity. 2005; 23(3):249-62. DOI: 10.1016/j.immuni.2005.08.001. View

20.

Bertrand F, Muller S, Roh K, Laurent C, Dupre L, Valitutti S . An initial and rapid step of lytic granule secretion precedes microtubule organizing center polarization at the cytotoxic T lymphocyte/target cell synapse. Proc Natl Acad Sci U S A. 2013; 110(15):6073-8. PMC: 3625254. DOI: 10.1073/pnas.1218640110. View