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

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<sup>+</sup> 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.

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Journal
Proceedings of the National Academy of Sciences
Published
2025-02-04
DOI
https://doi.org/10.1073/pnas.2413866122
Citations
5
Primary Topic
Angiogenesis and VEGF in Cancer
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article
Field-Weighted Citation Impact
4.03

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article

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

Nagaja Capitani, Nadia Alawar, Ute Becherer, Cosima T. Baldari et al.
5 citations
Proceedings of the National Academy of Sciences
Angiogenesis and VEGF in Cancer
4.03
article

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

Nagaja Capitani, Nadia Alawar, Ute Becherer, Cosima T. Baldari, Livio Trentin, Vanessa Tatangelo, Salvatore Valvo, Hsin‐Fang Chang, Ewoud B. Compeer, Claudia Schirra, Anna Onnis, Szu-Min Tu, Giuseppe Marotta, Michael Loran Dustin, Jemma Nicholls, Ludovica Lopresti, Bruce R. Blazar, Francesca Finetti, Chiara Cassioli, Laura Patrussi, Matthew J. A. Wood, Carmela Tangredi, Claire C. Staton, Annachiara Miccoli
article en
5 citations

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<sup>+</sup> 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.

Proceedings of the National Academy of SciencesVol. 122(6)
University of Siena (IT), University of Minnesota (US), University of Padua (IT), California Institute for Regenerative Medicine (US), University of Oxford (GB), Regenerative Medicine Institute (MX), University of Minnesota Medical Center (US), Cellular Therapeutics (United Kingdom) (GB), Saarland University (DE)
University of Minnesota, Università degli Studi di Siena, Kennedy Trust for Rheumatology Research, University of Oxford, European Commission, Università degli Studi di Padova, Universität des Saarlandes, Clarendon Fund, National Heart, Lung, and Blood Institute
Openalex Percentile: Top 7%
Angiogenesis and VEGF in Cancer
4.03
DOIOpen Access PDFOpenAlex
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