{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T15:25:56Z","timestamp":1791473156484,"version":"4.3.4"},"reference-count":50,"publisher":"BMJ","issue":"3","license":[{"start":{"date-parts":[[2022,3,10]],"date-time":"2022-03-10T00:00:00Z","timestamp":1646870400000},"content-version":"unspecified","delay-in-days":9,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"content-domain":{"domain":["bmj.com"],"crossmark-restriction":true},"short-container-title":["J Immunother Cancer"],"accepted":{"date-parts":[[2022,2,8]]},"published-print":{"date-parts":[[2022,3]]},"abstract":"<jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>CD73 is an ectonucleotidase producing the immunosuppressor mediator adenosine. Elevated levels of circulating CD73 in patients with cancer have been associated with disease progression and poor response to immunotherapy. Immunosuppressive pathways associated with exosomes can affect T-cell function and the therapeutic efficacy of anti-programmed cell-death protein 1 (anti-PD-1) therapy. Here, we conducted a retrospective pilot study to evaluate levels of exosomal CD73 before and early during treatment with anti-PD-1 agents in patients with melanoma and its potential contribution to affect T-cell functions and to influence the clinical outcomes of anti-PD-1 monotherapy.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>Exosomes were isolated by mini size exclusion chromatography from serum of patients with melanoma (n=41) receiving nivolumab or pembrolizumab monotherapy. Expression of CD73 and programmed death-ligand 1 (PD-L1) were evaluated on exosomes enriched for CD63 by on-bead flow cytometry. The CD73 AMPase activity was evaluated by mass spectrometry, also in the presence of selective inhibitors of CD73. Interferon (IFN)-\u03b3 production and granzyme B expression were measured in CD3\/28 activated T cells incubated with exosomes in presence of the CD73 substrate AMP. Levels of CD73 and PD-L1 on exosomes were correlated with therapy response. Exosomes isolated from healthy subjects were used as control.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>Isolated exosomes carried CD73 on their surface, which is enzymatically active in producing adenosine. Incubation of exosomes with CD3\/28 activated T cells in the presence of AMP resulted in a significant reduction of IFN-\u03b3 release, which was reversed by the CD73 inhibitor APCP or by the selective A2A adenosine receptor antagonist ZM241385. Expression levels of exosomal CD73 from serum of patients with melanoma were not significantly different from those in healthy subjects. Early on-treatment, expression levels of both CD73 and PD-L1 on exosomes isolated from patients receiving pembrolizumab or nivolumab monotherapy were significantly increased compared with baseline. Early during therapy exosomal PD-L1 increased in responders, while exosomal CD73 resulted significantly increased in non-responders.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>CD73 expressed on exosomes from serum of patients with melanoma produces adenosine and contributes to suppress T-cell functions. Early on-treatment, elevated expression levels of exosomal CD73 might affect the response to anti-PD-1 agents in patients with melanoma who failed to respond to therapy.<\/jats:p>\n               <\/jats:sec>","DOI":"10.1136\/jitc-2021-004043","type":"journal-article","created":{"date-parts":[[2022,3,10]],"date-time":"2022-03-10T11:45:37Z","timestamp":1646912737000},"page":"e004043","update-policy":"https:\/\/doi.org\/10.1136\/crossmarkpolicy","source":"Crossref","is-referenced-by-count":77,"title":["Exosomal CD73 from serum of patients with melanoma suppresses lymphocyte functions and is associated with therapy resistance to anti-PD-1 agents"],"prefix":"10.1136","volume":"10","author":[{"given":"Roberta","family":"Turiello","sequence":"first","affiliation":[{"name":"Department of Pharmacy, University of Salerno, Fisciano, Italy"},{"name":"PhD Program in Drug Discovery and Development, University of Salerno, Fisciano, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mariaelena","family":"Capone","sequence":"additional","affiliation":[{"name":"Istituto Nazionale Tumori IRCCS Fondazione G Pascale, Napoli, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elva","family":"Morretta","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, University of Salerno, Fisciano, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria Chiara","family":"Monti","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, University of Salerno, Fisciano, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9395-3190","authenticated-orcid":false,"given":"Gabriele","family":"Madonna","sequence":"additional","affiliation":[{"name":"Istituto Nazionale Tumori IRCCS Fondazione G Pascale, Napoli, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rosa","family":"Azzaro","sequence":"additional","affiliation":[{"name":"Istituto Nazionale Tumori IRCCS Fondazione G Pascale, Napoli, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pasquale","family":"Del Gaudio","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, University of Salerno, Fisciano, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ester","family":"Simeone","sequence":"additional","affiliation":[{"name":"Istituto Nazionale Tumori IRCCS Fondazione G Pascale, Napoli, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Antonio","family":"Sorrentino","sequence":"additional","affiliation":[{"name":"Istituto Nazionale Tumori IRCCS Fondazione G Pascale, Napoli, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paolo A","family":"Ascierto","sequence":"additional","affiliation":[{"name":"Istituto Nazionale Tumori IRCCS Fondazione G Pascale, Napoli, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6541-0997","authenticated-orcid":false,"given":"Silvana","family":"Morello","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, University of Salerno, Fisciano, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"239","published-online":{"date-parts":[[2022,3,10]]},"reference":[{"key":"2025100306180366000_10.3.e004043.1","doi-asserted-by":"crossref","first-page":"236","DOI":"10.1016\/j.semcancer.2019.08.002","article-title":"Unraveling the crosstalk between melanoma and immune cells in the tumor microenvironment","volume":"59","author":"Marzagalli","year":"2019","journal-title":"Semin Cancer Biol"},{"key":"2025100306180366000_10.3.e004043.2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(21)01206-X"},{"key":"2025100306180366000_10.3.e004043.3","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1910836"},{"key":"2025100306180366000_10.3.e004043.4","doi-asserted-by":"publisher","DOI":"10.1172\/JCI81135"},{"key":"2025100306180366000_10.3.e004043.5","doi-asserted-by":"publisher","DOI":"10.3747\/co.v16i3.367"},{"key":"2025100306180366000_10.3.e004043.6","doi-asserted-by":"publisher","DOI":"10.2174\/22115528113020020001"},{"key":"2025100306180366000_10.3.e004043.7","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0005219"},{"key":"2025100306180366000_10.3.e004043.8","doi-asserted-by":"publisher","DOI":"10.1038\/nm.2753"},{"key":"2025100306180366000_10.3.e004043.9","doi-asserted-by":"crossref","first-page":"2012","DOI":"10.3390\/jcm4121957","article-title":"Detection of exosomal miRNAs in the plasma of melanoma patients","volume":"4","author":"Pfeffer","year":"2015","journal-title":"J Clin Med"},{"key":"2025100306180366000_10.3.e004043.10","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-018-0392-8"},{"key":"2025100306180366000_10.3.e004043.11","doi-asserted-by":"publisher","DOI":"10.1038\/s41577-019-0264-y"},{"key":"2025100306180366000_10.3.e004043.12","doi-asserted-by":"crossref","first-page":"611","DOI":"10.1038\/s41571-020-0382-2","article-title":"The adenosine pathway in immuno-oncology","volume":"17","author":"Allard","year":"2020","journal-title":"Nat Rev Clin Oncol"},{"key":"2025100306180366000_10.3.e004043.13","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamcr.2008.01.024"},{"key":"2025100306180366000_10.3.e004043.14","doi-asserted-by":"publisher","DOI":"10.4161\/onci.26246"},{"key":"2025100306180366000_10.3.e004043.15","doi-asserted-by":"crossref","first-page":"6.1","DOI":"10.1146\/annurev-med-060619-023155","article-title":"Inhibition of the adenosine pathway to potentiate cancer immunotherapy: potential for combinatorial approaches","volume":"72","author":"Thompson","year":"2021","journal-title":"Annu Rev Med"},{"key":"2025100306180366000_10.3.e004043.16","doi-asserted-by":"publisher","DOI":"10.1200\/JCO.2018.36.15_suppl.4123"},{"key":"2025100306180366000_10.3.e004043.17","doi-asserted-by":"crossref","first-page":"S1320","DOI":"10.1016\/j.annonc.2021.08.2121","article-title":"LBA42 coast: an open-label, randomised, phase II platform study of durvalumab alone or in combination with novel agents in patients with locally advanced, unresectable, stage III NSCLC","volume":"32","author":"Martinez-Marti","year":"2021","journal-title":"Annals of Oncology"},{"key":"2025100306180366000_10.3.e004043.18","doi-asserted-by":"crossref","DOI":"10.3389\/fphar.2020.609931","article-title":"Cd73: a promising biomarker in cancer patients","volume":"11","author":"Turiello","year":"2020","journal-title":"Front Pharmacol"},{"key":"2025100306180366000_10.3.e004043.19","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2019.00010"},{"key":"2025100306180366000_10.3.e004043.20","doi-asserted-by":"publisher","DOI":"10.1186\/s12967-017-1348-8"},{"key":"2025100306180366000_10.3.e004043.21","doi-asserted-by":"publisher","DOI":"10.1136\/jitc-2020-001689"},{"key":"2025100306180366000_10.3.e004043.22","doi-asserted-by":"publisher","DOI":"10.1016\/j.jim.2014.06.007"},{"key":"2025100306180366000_10.3.e004043.23","doi-asserted-by":"publisher","DOI":"10.1111\/cei.12354"},{"key":"2025100306180366000_10.3.e004043.24","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1111\/cei.13157","article-title":"Exosomes in HNSCC plasma as surrogate markers of tumour progression and immune competence","volume":"194","author":"Theodoraki","year":"2018","journal-title":"Clin Exp Immunol"},{"key":"2025100306180366000_10.3.e004043.25","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1003884"},{"key":"2025100306180366000_10.3.e004043.26","doi-asserted-by":"publisher","DOI":"10.1002\/eji.201242909"},{"key":"2025100306180366000_10.3.e004043.27","doi-asserted-by":"crossref","first-page":"738","DOI":"10.1016\/j.immuni.2019.01.010","article-title":"Specific decrease in B-cell-derived extracellular vesicles enhances post-chemotherapeutic CD8+ T cell responses","volume":"50","author":"Zhang","year":"2019","journal-title":"Immunity"},{"key":"2025100306180366000_10.3.e004043.28","doi-asserted-by":"crossref","first-page":"599","DOI":"10.1007\/s10456-020-09728-8","article-title":"Tumor-derived exosomes promote angiogenesis via adenosine A\n                  2B\n                receptor signaling","volume":"23","author":"Ludwig","year":"2020","journal-title":"Angiogenesis"},{"key":"2025100306180366000_10.3.e004043.29","doi-asserted-by":"crossref","DOI":"10.1038\/s41598-019-56542-4","article-title":"Melanoma cell-derived exosomes in plasma of melanoma patients suppress functions of immune effector cells","volume":"10","author":"Sharma","year":"2020","journal-title":"Sci Rep"},{"key":"2025100306180366000_10.3.e004043.30","doi-asserted-by":"crossref","DOI":"10.1038\/s41467-021-26134-w","article-title":"CD73-mediated adenosine production by CD8 T cell-derived extracellular vesicles constitutes an intrinsic mechanism of immune suppression","volume":"12","author":"Schneider","year":"2021","journal-title":"Nat Commun"},{"key":"2025100306180366000_10.3.e004043.31","doi-asserted-by":"publisher","DOI":"10.1158\/1078-0432.CCR-16-2819"},{"key":"2025100306180366000_10.3.e004043.32","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1007\/978-1-4939-7142-8_16","article-title":"Isolation of biologically active exosomes from plasma of patients with cancer","volume":"1633","author":"Hong","year":"2017","journal-title":"Methods Mol Biol"},{"key":"2025100306180366000_10.3.e004043.33","doi-asserted-by":"crossref","first-page":"1435138","DOI":"10.1080\/20013078.2018.1435138","article-title":"Immunoaffinity-Based isolation of melanoma cell-derived exosomes from plasma of patients with melanoma","volume":"7","author":"Sharma","year":"2018","journal-title":"J Extracell Vesicles"},{"key":"2025100306180366000_10.3.e004043.34","doi-asserted-by":"crossref","first-page":"1692401","DOI":"10.1080\/20013078.2019.1692401","article-title":"Purity and yield of melanoma exosomes are dependent on isolation method","volume":"9","author":"Shu","year":"2020","journal-title":"J Extracell Vesicles"},{"key":"2025100306180366000_10.3.e004043.35","doi-asserted-by":"publisher","DOI":"10.1038\/nri855"},{"key":"2025100306180366000_10.3.e004043.36","doi-asserted-by":"publisher","DOI":"10.1038\/nrc.2017.86"},{"key":"2025100306180366000_10.3.e004043.37","doi-asserted-by":"crossref","first-page":"162","DOI":"10.3389\/fimmu.2019.00162","article-title":"Adenosine A2A receptor stimulation inhibits TCR-induced Notch1 activation in CD8+T-Cells","volume":"10","author":"Sorrentino","year":"2019","journal-title":"Front Immunol"},{"key":"2025100306180366000_10.3.e004043.38","doi-asserted-by":"crossref","first-page":"927","DOI":"10.1038\/s41416-021-01413-x","article-title":"Cancer immunotherapy: it's time to better predict patients' response","volume":"125","author":"Pilard","year":"2021","journal-title":"Br J Cancer"},{"key":"2025100306180366000_10.3.e004043.39","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2017.01.017"},{"key":"2025100306180366000_10.3.e004043.40","doi-asserted-by":"publisher","DOI":"10.1097\/PPO.0000000000000303"},{"key":"2025100306180366000_10.3.e004043.41","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0605251103"},{"key":"2025100306180366000_10.3.e004043.42","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2019.00925"},{"key":"2025100306180366000_10.3.e004043.43","doi-asserted-by":"crossref","DOI":"10.3390\/cancers13123034","article-title":"Soluble programmed death ligand-1 (sPD-L1): a pool of circulating proteins implicated in health and diseases","volume":"13","author":"Bailly","year":"2021","journal-title":"Cancers"},{"key":"2025100306180366000_10.3.e004043.44","doi-asserted-by":"publisher","DOI":"10.1016\/j.ccell.2020.08.005"},{"key":"2025100306180366000_10.3.e004043.45","doi-asserted-by":"crossref","first-page":"1189","DOI":"10.1016\/j.celrep.2017.04.031","article-title":"Interferon receptor signaling pathways regulating PD-L1 and PD-L2 expression","volume":"19","author":"Garcia-Diaz","year":"2017","journal-title":"Cell Rep"},{"key":"2025100306180366000_10.3.e004043.46","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2019.02.016"},{"key":"2025100306180366000_10.3.e004043.47","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1604958"},{"key":"2025100306180366000_10.3.e004043.48","doi-asserted-by":"crossref","first-page":"4509","DOI":"10.1002\/cam4.1700","article-title":"Interferon gamma in cancer immunotherapy","volume":"7","author":"Ni","year":"2018","journal-title":"Cancer Med"},{"key":"2025100306180366000_10.3.e004043.49","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-17-0395"},{"key":"2025100306180366000_10.3.e004043.50","doi-asserted-by":"publisher","DOI":"10.1038\/nrc3498"}],"updated-by":[{"DOI":"10.1136\/jitc-2021-004043corr1","type":"correction","label":"Correction","source":"publisher","updated":{"date-parts":[[2023,7,18]],"date-time":"2023-07-18T00:00:00Z","timestamp":1689638400000}}],"container-title":["Journal for ImmunoTherapy of Cancer"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1136\/jitc-2021-004043","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,3]],"date-time":"2025-10-03T13:18:16Z","timestamp":1759497496000},"score":1,"resource":{"primary":{"URL":"https:\/\/jitc.bmj.com\/lookup\/doi\/10.1136\/jitc-2021-004043"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,3]]},"references-count":50,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2022,3,10]]},"published-print":{"date-parts":[[2022,3]]}},"alternative-id":["10.1136\/jitc-2021-004043"],"URL":"https:\/\/doi.org\/10.1136\/jitc-2021-004043","relation":{"correction":[{"id-type":"doi","id":"10.1136\/jitc-2021-004043corr1","asserted-by":"object"}]},"ISSN":["2051-1426"],"issn-type":[{"value":"2051-1426","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,3]]}}}