SARA ANDREONE

PhD Graduate

PhD program:: XXXV


supervisor: Giovanna Schiavoni
co-supervisor: Silvia Piconese

Thesis title: Eosinophils acquire immune checkpoint molecules through trogocytosis: implications in cancer immunity.

Trogocytosis is a cellular process whereby a cell acquires a membrane fragment from a donor cell in a contact-dependent manner allowing for transfer of surface proteins with functional integrity. Trogocytosis has been described in most immune cells, with the exception of eosinophils, and represents an important mechanism for the regulation of anti-tumor immune responses. Eosinophils play important roles in cancer immunity through release of soluble mediators and contact-dependent mechanisms. We previously reported that activation of eosinophils with the alarmin IL-33 promotes adhesion to cancer cells via the CD11b/CD18 integrin complex resulting in tumor cell killing. Here, we evaluated trogocytosis in eosinophils following activation with IL-33 and upon contact with tumor cells. Flow cytometry and confocal microscopy demonstrated that after 1-hour co-culture with tumor cells (EG.7-OVA, MC38, TC-1 and B16.F10) labelled with a membrane dye, IL-33 activated, but not resting, eosinophils acquire membrane fragments from target cells. Time-lapse video microscopy revealed that trogocytosis by activated eosinophils occurs rapidly, within 2 minutes after contact with the target tumor cell. Transmission electron microscopy (TEM) showed trogocytic invaginations at the interface between activated eosinophils and tumor cells. Blockade of CD11b/CD18 on eosinophils membrane significantly reduced trogocytosis, highlighting the role of this integrin complex for binding to target tumor cells and formation of an immunological synapse. Since in immune cell trogocytosis is typically mediated by receptor–ligand interactions, we studied the ability of eosinophils to acquire specific proteins expressed by tumor cells, but not by eosinophils, focusing on immune checkpoint molecules. Eosinophils do not express certain immune checkpoints, such as PD-1 and TIGIT, but acquired these molecules after 1-hour co-culture with PD-1/TIGIT-expressing EG.7-OVA lymphoma cells. In contrast, eosinophils expressed PD-L1 that was further up-regulated by IL-33. Blocking the PD-1/PD-L1 interaction with αPD-L1 antibody markedly reduced PD-1 trogocytosis, demonstrating that the acquisition of PD-1 operated by eosinophils is receptor-dependent. Furthermore, blocking the eosinophil-tumor interaction of CD11b/CD18 also reduced PD-1 trogocytosis, denoting the requirement of cell-cell contact for this process. Interestingly, stimuli capable of up-regulating either CD11b/CD18 (i.e., CCL11) or PD-L1 (i.e., LPS), were not sufficient in triggering PD-1 trogocytosis in eosinophils, suggesting the need for induction of both signals for this process to occur. Finally, we provide a model of immunological synapse that may occur during eosinophils trogocytosis. Overall, our findings demonstrate that eosinophils can acquire tumor-expressed molecules through trogocytosis and that this process is strongly enhanced by activation stimuli (i.e., IL-33) that promote CD11b/CD18-dependent cell adhesion and increase the expression of specific ligands. The acquisition of immune checkpoints by eosinophils in the tumor microenvironment may potentially affect immunotherapy response in cancer patients.

Research products

11573/1656116 - 2022 - Trogocytosis in innate immunity to cancer is an intimate relationship with unexpected outcomes
Mattei, Fabrizio; Andreone, Sara; Spadaro, Francesca; Noto, Francesco; Tinari, Antonella; Falchi, Mario; Piconese, Silvia; Afferni, Claudia; Schiavoni, Giovanna - 01a Articolo in rivista
paper: ISCIENCE ([Cambridge MA] : Cell Press Elsevier Inc.) pp. - - issn: 2589-0042 - wos: WOS:000869007300005 (4) - scopus: 2-s2.0-85138400626 (6)

11573/1549840 - 2021 - Anticancer effects of sublingual type I IFN in combination with chemotherapy in implantable and spontaneous tumor models
Ciccolella, M.; Andreone, S.; Mancini, J.; Sestili, P.; Negri, D.; Pacca, A. M.; D'urso, M. T.; Macchia, D.; Canese, R.; Pang, K.; Saiying Ko, T.; Decadt, Y.; Schiavoni, G.; Mattei, F.; Belardelli, F.; Arico, E.; Bracci, L. - 01a Articolo in rivista
paper: CELLS (Basel: mdpi-Molecular Diversity Preservation International) pp. - - issn: 2073-4409 - wos: WOS:000642862800001 (4) - scopus: 2-s2.0-85105143158 (4)

11573/1549845 - 2021 - Oncoimmunology meets organs-on-chip
Mattei, F.; Andreone, S.; Mencattini, A.; De Ninno, A.; Businaro, L.; Martinelli, E.; Schiavoni, G. - 01g Articolo di rassegna (Review)
paper: FRONTIERS IN MOLECULAR BIOSCIENCES (Lausanne : Frontiers Media S.A., 2014-) pp. - - issn: 2296-889X - wos: WOS:000638198400001 (25) - scopus: 2-s2.0-85103873918 (22)

11573/1529300 - 2021 - A Clonogenic Assay to Quantify Melanoma Micrometastases in Pulmonary Tissue
Mattei, F.; Andreone, S.; Schiavoni, G. - 02a Capitolo o Articolo
book: Methods in Molecular Biology - (978-1-0716-1204-0; 978-1-0716-1205-7)

11573/1549837 - 2021 - LXR directly regulates glycosphingolipid synthesis and affects human CD4+ T cell function
Waddington, Kirsty E; Robinson, George A; Rubio-Cuesta, Beatriz; Chrifi-Alaoui, Eden; Andreone, Sara; Poon, Kok-Siong; Ivanova, Iveta; Martin-Gutierrez, Lucia; Owen, Dylan M; Jury, Elizabeth C; Pineda-Torra, Inés - 01a Articolo in rivista
paper: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA (National Academy of Sciences:2101 Constitution Avenue Northwest:Washington, DC 20418:(877)314-2253, (615)377-3322, EMAIL: subspnas@nas.edu, INTERNET: http://www.pnas.org, Fax: (615)377-0525) pp. - - issn: 0027-8424 - wos: WOS:000659439900018 (23) - scopus: 2-s2.0-85106150387 (26)

11573/1478777 - 2020 - Anti-tumorigenic activities of IL-33: a mechanistic insight
Andreone, S.; Gambardella, A. R.; Mancini, J.; Loffredo, S.; Marcella, S.; La Sorsa, V.; Varricchi, G.; Schiavoni, G.; Mattei, F. - 01g Articolo di rassegna (Review)
paper: FRONTIERS IN IMMUNOLOGY (Lausanne : Frontiers Research Foundation, 2010-) pp. - - issn: 1664-3224 - wos: WOS:000598134800001 (23) - scopus: 2-s2.0-85097614947 (22)

11573/1455658 - 2020 - Eosinophils in the Tumor Microenvironment
Mattei, F.; Andreone, S.; Marone, G.; Gambardella, A. R.; Loffredo, S.; Varricchi, G.; Schiavoni, G. - 02a Capitolo o Articolo
book: Advances in Experimental Medicine and Biology - (978-3-030-49269-4; 978-3-030-49270-0)

11573/1342552 - 2019 - IL-33 promotes CD11b/CD18-mediated adhesion of eosinophils to cancer cells and synapse-polarized degranulation leading to tumor cell killing
Andreone, S.; Spadaro, F.; Buccione, C.; Mancini, J.; Tinari, A.; Sestili, P.; Gambardella, A. R.; Lucarini, V.; Ziccheddu, G.; Parolini, I.; Zanetti, C.; D'urso, M. T.; De Ninno, A.; Businaro, L.; Afferni, C.; Mattei, F.; Schiavoni, G. - 01a Articolo in rivista
paper: CANCERS (Basel: MDPI) pp. - - issn: 2072-6694 - wos: WOS:000502290100046 (48) - scopus: 2-s2.0-85074467395 (54)

11573/1342554 - 2018 - The pleiotropic immunomodulatory functions of IL-33 and its implications in tumor immunity
Afferni, C.; Buccione, C.; Andreone, S.; Galdiero, M. R.; Varricchi, G.; Marone, G.; Mattei, F.; Schiavoni, G. - 01g Articolo di rassegna (Review)
paper: FRONTIERS IN IMMUNOLOGY (Lausanne : Frontiers Research Foundation, 2010-) pp. - - issn: 1664-3224 - wos: WOS:000450023500003 (78) - scopus: 2-s2.0-85056663985 (83)

11573/1342558 - 2017 - IL-33 restricts tumor growth and inhibits pulmonary metastasis in melanoma-bearing mice through eosinophils
Lucarini, V.; Ziccheddu, G.; Macchia, I.; La Sorsa, V.; Peschiaroli, F.; Buccione, C.; Sistigu, A.; Sanchez, M.; Andreone, S.; D'urso, M. T.; Spada, M.; Macchia, D.; Afferni, C.; Mattei, F.; Schiavoni, G. - 01a Articolo in rivista
paper: ONCOIMMUNOLOGY (Austin, TX : Landes Bioscience) pp. - - issn: 2162-4011 - wos: WOS:000404014300009 (141) - scopus: 2-s2.0-85019598855 (149)

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