Activation-induced thrombospondin-4 works with thrombospondin-1 to build cytotoxic supramolecular attack particles
Abstract
Cytotoxic attack particles released by CTLs and NK cells include diverse phospholipid membrane and glycoprotein encapsulated entities that contribute to target cell killing. Supramolecular attack particles (SMAPs) are one type of particle characterized by a cytotoxic core enriched in granzymes and perforin surrounded by a proteinaceous shell including thrombospondin (TSP)-1. TSP-4 was also detected in bulk analysis of SMAPs released by CTLs; however, it has not been investigated whether TSP-4 contributes to distinct SMAP types or the same SMAP type as TSP-1 and, if in the same type of SMAP, whether TSP-4 and TSP-1 cooperate or compete. Here, we observed that TSP-4 expression increased upon CD8 + T cell activation while, surprisingly, TSP-1 was down-regulated. Correlative Light and Electron Microscopy and Stimulated Emission Depletion microscopy localized TSP-4 and TSP-1 in SMAP-containing multicore granules. Superresolution dSTORM revealed that TSP-4 and TSP-1 are usually enriched in the same SMAPs while particles with single-positive shells are rare. Retention Using Selective Hooks assays showed that TSP-4 localizes to the lytic granules faster than TSP-1 and promotes its accumulation therein. TSP-4 contributed to direct CTL-mediated killing, as previously shown for TSP-1. TSP-4 and TSP-1 were both required for latent SMAP-mediated cell killing, in which released SMAPs kill targets after removal of the CTLs. Of note, we found that chronic lymphocytic leukemia (CLL) cell culture supernatants suppressed expression of TSP-4 in CTL and latent SMAP-mediated killing. These results identify TSP-4 as a functionally important component of SMAPs and suggest that SMAPs may be targeted for immune suppression by CLL.
Article Details
Journal Info
Proceedings of the National Academy of Sciences
National Academy of Sciences
Authors (24)
Chiara Cassioli
Department of Life Sciences, University of Siena
Nagaja Capitani
Department of Life Sciences, University of Siena
Claire C. Staton
Kennedy Institute of Rheumatology, Nuffield Department of Orthopaedics, Rheumatology and Musculoskeletal Sciences, University of Oxford
Claudia Schirra
Department of Cellular Neurophysiology, Center for Integrative Physiology and Molecular Medicine, Saarland University
Francesca Finetti
Department of Life Sciences, University of Siena
Anna Onnis
Department of Life Sciences, University of Siena
Nadia Alawar
Department of Cellular Neurophysiology, Center for Integrative Physiology and Molecular Medicine, Saarland University
Szu-Min Tu
Department of Cellular Neurophysiology, Center for Integrative Physiology and Molecular Medicine, Saarland University
Ludovica Lopresti
Department of Life Sciences, University of Siena
Vanessa Tatangelo
Department of Life Sciences, University of Siena
Carmela Tangredi
Department of Life Sciences, University of Siena
Salvatore Valvo
The Kennedy Institute of Rheumatology, Nuffield Department of Orthopaedics, Rheumatology and Musculoskeletal Sciences (NDORMS), University of Oxford, Roosevelt Drive
Hsin-Fang Chang
Department of Cellular Neurophysiology, Center for Integrative Physiology and Molecular Medicine, Saarland University
Annachiara Miccoli
Department of Life Sciences, University of Siena
Ewoud B. Compeer
Kennedy Institute of Rheumatology, Nuffield Department of Orthopaedics, Rheumatology and Musculoskeletal Sciences, University of Oxford
Jemma Nicholls
Department of Pediatrics, Division of Blood & Marrow Transplant & Cellular Therapy, University of Minnesota
Bruce R. Blazar
Department of Pediatrics, Division of Blood & Marrow Transplant & Cellular Therapy, University of Minnesota
Giuseppe Marotta
Stem Cell Transplant and Cellular Therapy Unit, Siena University Hospital
Matthew J. A. Wood
Institute for Developmental and Regenerative Medicine, Department of Paediatrics, University of Oxford
Livio Trentin
Hematology Unit, Department of Medicine, University of Padua
Laura Patrussi
Department of Life Sciences, University of Siena
Michael L. Dustin
Ute Becherer
Department of Cellular Neurophysiology, Center for Integrative Physiology and Molecular Medicine, Saarland University
Cosima T. Baldari
Department of Life Sciences, University of Siena