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This study will investigate the clinical relevance of the tumor microenvironment in a large cohort of treatment\u2010na\u00efve patients affected by stage II\u2013IV LSCC.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Whole slide\u2010based digital pathology analysis was applied to measure six immune cell populations identified by immunohistochemistry (IHC) staining for CD3, CD8, CD20, CD66b, CD163 and CD38. Survival analysis was performed by Cox proportional hazards models and unsupervised hierarchical clustering using the k\u2010means method. Double IHC staining and <jats:italic>in\u2010situ<\/jats:italic> hybridisation by RNAscope allowed further analysis of a protumoral B cell population.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>A cohort of 98 patients was enrolled and analysed. The cluster of immune\u2010infiltrated LSCCs demonstrated a significantly worse disease\u2010specific survival rate. We also discovered a new association between high CD20<jats:sup>+<\/jats:sup> B cells and a greater risk of\u00a0distant recurrence. The phenotypic analysis of infiltrating CD20<jats:sup>+<\/jats:sup> B cells showed a na\u00efve (BCL6<jats:sup>\u2212<\/jats:sup>CD27<jats:sup>\u2212<\/jats:sup>Mum1<jats:sup>\u2212<\/jats:sup>) regulatory phenotype, producing TGF\u03b2 but not IL10, according to an active TGF\u03b2 pathway, as proved by positive pSMAD2 staining.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>The identification of regulatory B\u00a0cells in the context of LSCC, along with the activation of the TGF\u03b2 pathway, could provide the basis for new trials investigating the efficacy of already available molecules targeting the TGF\u03b2 pathway in the treatment of LSCC.<\/jats:p><\/jats:sec>","DOI":"10.1002\/cti2.1445","type":"journal-article","created":{"date-parts":[[2023,4,26]],"date-time":"2023-04-26T11:07:11Z","timestamp":1682507231000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["The prometastatic 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