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Tertiary lymphoid structures (TLS), as organized ectopic lymphoid aggregates, play a pivotal role in modulating antitumor immune responses. However, their landscape, clinical significance, and molecular determinants in osteosarcoma remain largely unexplored.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>TLS were identified in osteosarcoma tissues by immunohistochemistry and multiplex immunofluorescence. Using transcriptomic data from the TARGET, GEO (GSE21257, GSE16091, GSE39055), and PKUPH cohorts, we developed a TLS-based prognostic index (TLSPI) through integrative machine-learning modeling. Functional enrichment, immune infiltration, and immunotherapy response prediction analyses were performed to characterize TLSPI-related biological features. The role of the key TLS-associated gene CXCL13 was validated through immunohistochemistry, single-cell transcriptomic profiling, and in vitro functional experiments.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      TLS were detected in osteosarcoma and were significantly associated with improved overall survival (\n                      <jats:italic>p<\/jats:italic>\n                      \u2009=\u20090.024). Differential expression and enrichment analyses revealed that TLSPI-related genes were primarily involved in immune activation and antigen presentation pathways. Immune profiling revealed that the low-TLSPI group was characterized by greater immune infiltration and higher expression of immune checkpoint molecules, as well as lower TIDE scores, which indicated greater predicted sensitivity to immunotherapy. CXCL13 was identified as a key TLS-associated gene whose high expression correlated with improved survival. Immunohistochemical analysis confirmed the prognostic significance of CXCL13, while single-cell and functional assays demonstrated its association with B-cell activation within the osteosarcoma microenvironment.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>This study provides a comprehensive characterization of TLS in osteosarcoma and establishes a TLS-based prognostic index with potential clinical applicability. CXCL13 may serve as a critical mediator linking TLS formation and antitumor immunity, providing potential therapeutic implications for osteosarcoma immunotherapy.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1007\/s13402-026-01226-1","type":"journal-article","created":{"date-parts":[[2026,5,16]],"date-time":"2026-05-16T17:03:44Z","timestamp":1778951024000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Machine learning\u2013based integrative analysis identifies CXCL13-driven tertiary lymphoid structures as favorable immune and prognostic features in osteosarcoma"],"prefix":"10.1007","volume":"49","author":[{"given":"Jie","family":"Jiang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiuhui","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lu","family":"Xie","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yiyang","family":"Yu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xin","family":"Sun","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaojiao","family":"Sun","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shen","family":"Yang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huanmin","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tingting","family":"Ren","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaodong","family":"Tang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,5,16]]},"reference":[{"key":"1226_CR1","doi-asserted-by":"publisher","first-page":"609","DOI":"10.1038\/s41571-021-00519-8","volume":"18","author":"J Gill","year":"2021","unstructured":"J. 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