{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,6]],"date-time":"2025-11-06T12:26:15Z","timestamp":1762431975446,"version":"build-2065373602"},"reference-count":36,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2021,1,23]],"date-time":"2021-01-23T00:00:00Z","timestamp":1611360000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"FEDER - Fundo Europeu de Desenvolvimento Regional funds through the COMPETE 2020 - Operacional Program for Competitiveness and Internationalisation (POCI), PORTUGAL 2020, Portuguese funds through Portuguese Science &amp; Technology Foundation (FCT) \/ Minist\u00e9r","award":["\"Institute for Research and Innovation in Health Sciences\" (POCI-01-0145-FEDER-007274)"],"award-info":[{"award-number":["\"Institute for Research and Innovation in Health Sciences\" (POCI-01-0145-FEDER-007274)"]}]},{"name":"Norte Portugal Regional Program (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF","award":["Project CANCER Ref. NORTE-01-0145-FEDER-000029 and DOCnet Ref. NORTE-01-0145-FEDER-000003)"],"award-info":[{"award-number":["Project CANCER Ref. NORTE-01-0145-FEDER-000029 and DOCnet Ref. NORTE-01-0145-FEDER-000003)"]}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["Project Ref. PTDC\/BBB-ECT\/2518\/2014","FCT fellowship PD\/BD\/128406\/2017 to SR, and Junior Research contract to JC (regulated by the decree-law 57\/2016 emended by law 57\/2017)","IPATIMUP integrates the i3S Research Unit, which is partially supported by FCT"],"award-info":[{"award-number":["Project Ref. PTDC\/BBB-ECT\/2518\/2014","FCT fellowship PD\/BD\/128406\/2017 to SR, and Junior Research contract to JC (regulated by the decree-law 57\/2016 emended by law 57\/2017)","IPATIMUP integrates the i3S Research Unit, which is partially supported by FCT"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"name":"FCT PhD Programs and by Programa Operacional Potencial Humano (POCH)","award":["BiotechHealth Programe (Doctoral Program on Cellular and Molecular Biotechnology Applied to Health Sciences"],"award-info":[{"award-number":["BiotechHealth Programe (Doctoral Program on Cellular and Molecular Biotechnology Applied to Health Sciences"]}]},{"name":"IPATIMUP Board of Directors","award":["\"The role of gastric cancer cell-derived extracellular vesicles\u201d"],"award-info":[{"award-number":["\"The role of gastric cancer cell-derived extracellular vesicles\u201d"]}]},{"name":"COMPETE 2020 - POCI, Lisboa Portugal Regional Operational Program (Lisboa2020), Algarve Portugal Regional Operational Program (CRESC Algarve2020), under the PORTUGAL 2020 Partnership Agreement, through ERDF, and by FCT","award":["GenomePT project (POCI-01-0145-FEDER-022184)"],"award-info":[{"award-number":["GenomePT project (POCI-01-0145-FEDER-022184)"]}]},{"name":"European Union\u2019s Horizon 2020 research and innovation program","award":["Marie Sk\u0142odowska-Curie grant agreement No. 722148 to Train-EV project"],"award-info":[{"award-number":["Marie Sk\u0142odowska-Curie grant agreement No. 722148 to Train-EV project"]}]},{"name":"ERDF, POCI and FCT","award":["Project CANCERSTEM"],"award-info":[{"award-number":["Project CANCERSTEM"]}]},{"DOI":"10.13039\/100010663","name":"H2020 European Research Council","doi-asserted-by":"publisher","award":["NFCCdM has received funding from the European Research Council (ERC) under the European Union\u2019s Horizon 2020 research and innovation program (grant agreement No. 852832)."],"award-info":[{"award-number":["NFCCdM has received funding from the European Research Council (ERC) under the European Union\u2019s Horizon 2020 research and innovation program (grant agreement No. 852832)."]}],"id":[{"id":"10.13039\/100010663","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cancers"],"abstract":"<jats:p>Gastric cancer (GC) patients display increased regulatory T cell (Tregs) numbers in peripheral blood and among tumor-infiltrating lymphocytes. Nevertheless, the role of Tregs in GC progression remains controversial. Here, we sought to explore the impact of Tregs in GCs with distinct histology, and whether Tregs can directly influence tumor cell behavior and GC development. We performed a comprehensive immunophenotyping of 82 human GC cases, through an integrated analysis of multispectral immunofluorescence detection of T cells markers and patient clinicopathological data. Moreover, we developed 3D in vitro co-cultures with Tregs and tumor cells that were followed by high-throughput and light-sheet imaging, and their biological features studied with conventional\/imaging flow cytometry and Western blotting. We showed that Tregs located at the tumor nest were frequent in intestinal-type GCs but did not associate with increased levels of effector T cells. Our in vitro results suggested that Tregs preferentially infiltrated intestinal-type GC spheroids, induced the expression of IL2R\u03b1 and activation of MAPK signaling pathway in tumor cells, and promoted spheroid growth. Accumulation of Tregs in intestinal-type GCs was increased at early stages of the stomach wall invasion and in the absence of vascular and perineural invasion. In this study, we proposed a non-immunosuppressive mechanism through which Tregs might directly modulate GC cells and thereby promote tumor growth. Our findings hold insightful implications for therapeutic strategies targeting intestinal-type GCs and other tumors with similar immune context.<\/jats:p>","DOI":"10.3390\/cancers13030421","type":"journal-article","created":{"date-parts":[[2021,1,25]],"date-time":"2021-01-25T02:07:00Z","timestamp":1611540420000},"page":"421","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Immunophenotype of Gastric Tumors Unveils a Pleiotropic Role of Regulatory T Cells in Tumor Development"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5390-3807","authenticated-orcid":false,"given":"Sara","family":"Rocha","sequence":"first","affiliation":[{"name":"i3S\u2014Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, Universidade do Porto, 4200-135 Porto, Portugal"},{"name":"Ipatimup\u2014Institute of Molecular Pathology and Immunology of University of Porto, 4200-135 Porto, Portugal"},{"name":"Doctoral Program on Cellular and Molecular Biotechnology Applied to Health Sciences, ICBAS\u2014Instituto de Ci\u00eancias Biom\u00e9dicas Abel Salazar, Universidade do Porto, 4050-313 Porto, Portugal"}]},{"given":"Afonso P","family":"Basto","sequence":"additional","affiliation":[{"name":"iMM\u2014Instituto de Medicina Molecular Jo\u00e3o Lobo Antunes, Faculdade de Medicina da Universidade de Lisboa, 1649-028 Lisbon, Portugal"},{"name":"Instituto Gulbenkian de Ci\u00eancia, 2780-156 Oeiras, Portugal"}]},{"given":"Marieke E","family":"Ijsselsteijn","sequence":"additional","affiliation":[{"name":"Department of Pathology, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7409-9651","authenticated-orcid":false,"given":"Sara P","family":"Teles","sequence":"additional","affiliation":[{"name":"i3S\u2014Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, Universidade do Porto, 4200-135 Porto, Portugal"},{"name":"Ipatimup\u2014Institute of Molecular Pathology and Immunology of University of Porto, 4200-135 Porto, Portugal"}]},{"given":"Maria M","family":"Azevedo","sequence":"additional","affiliation":[{"name":"i3S\u2014Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, Universidade do Porto, 4200-135 Porto, Portugal"}]},{"given":"Gilza","family":"Gon\u00e7alves","sequence":"additional","affiliation":[{"name":"Department of Pathology, Faculty of Medicine of the University of Porto (FMUP), 4200-319 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3377-0530","authenticated-orcid":false,"given":"Irene","family":"Gullo","sequence":"additional","affiliation":[{"name":"i3S\u2014Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, Universidade do Porto, 4200-135 Porto, Portugal"},{"name":"Ipatimup\u2014Institute of Molecular Pathology and Immunology of University of Porto, 4200-135 Porto, Portugal"},{"name":"Department of Pathology, Faculty of Medicine of the University of Porto (FMUP), 4200-319 Porto, Portugal"},{"name":"Department of Pathology, Centro Hospitalar Universit\u00e1rio de S\u00e3o Jo\u00e3o, 4200-319 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8712-7394","authenticated-orcid":false,"given":"Gabriela M","family":"Almeida","sequence":"additional","affiliation":[{"name":"i3S\u2014Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, Universidade do Porto, 4200-135 Porto, Portugal"},{"name":"Ipatimup\u2014Institute of Molecular Pathology and Immunology of University of Porto, 4200-135 Porto, Portugal"},{"name":"Department of Pathology, Faculty of Medicine of the University of Porto (FMUP), 4200-319 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0800-4167","authenticated-orcid":false,"given":"Joaqu\u00edn J","family":"Maqueda","sequence":"additional","affiliation":[{"name":"i3S\u2014Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, Universidade do Porto, 4200-135 Porto, Portugal"},{"name":"Ipatimup\u2014Institute of Molecular Pathology and Immunology of University of Porto, 4200-135 Porto, Portugal"}]},{"given":"Marta I","family":"Oliveira","sequence":"additional","affiliation":[{"name":"International Iberian Nanotechnology Laboratory, 4715-330 Braga, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1964-1006","authenticated-orcid":false,"given":"F\u00e1tima","family":"Carneiro","sequence":"additional","affiliation":[{"name":"i3S\u2014Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, Universidade do Porto, 4200-135 Porto, Portugal"},{"name":"Ipatimup\u2014Institute of Molecular Pathology and Immunology of University of Porto, 4200-135 Porto, Portugal"},{"name":"Department of Pathology, Faculty of Medicine of the University of Porto (FMUP), 4200-319 Porto, Portugal"},{"name":"Department of Pathology, Centro Hospitalar Universit\u00e1rio de S\u00e3o Jo\u00e3o, 4200-319 Porto, Portugal"}]},{"given":"Jo\u00e3o T","family":"Barata","sequence":"additional","affiliation":[{"name":"iMM\u2014Instituto de Medicina Molecular Jo\u00e3o Lobo Antunes, Faculdade de Medicina da Universidade de Lisboa, 1649-028 Lisbon, Portugal"}]},{"given":"Lu\u00eds","family":"Gra\u00e7a","sequence":"additional","affiliation":[{"name":"iMM\u2014Instituto de Medicina Molecular Jo\u00e3o Lobo Antunes, Faculdade de Medicina da Universidade de Lisboa, 1649-028 Lisbon, Portugal"},{"name":"Instituto Gulbenkian de Ci\u00eancia, 2780-156 Oeiras, Portugal"}]},{"given":"Noel F C C","family":"de Miranda","sequence":"additional","affiliation":[{"name":"Department of Pathology, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands"}]},{"given":"Joana","family":"Carvalho","sequence":"additional","affiliation":[{"name":"i3S\u2014Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, Universidade do Porto, 4200-135 Porto, Portugal"},{"name":"Ipatimup\u2014Institute of 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