Supplementary file 1_Minimally expanded breast cancer tumor-infiltrating-lymphocytes provide guidance for therapeutic selection.docx
Introduction<p>The analysis of tumor-infiltrating lymphocytes often requires techniques that expand their numbers, potentially introducing bias. To address this, we performed a detailed analysis of minimally cultured TILs to evaluate whether this approach better preserves their characteristics...
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2025
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| author | Andrea Aran (11711939) |
| author2 | Gonzalo Lázaro (16752705) Vicente Marco (6068645) Vicente Peg (466433) Maitane Faus (22686683) Laia Garrigós (11644057) José Pérez-García (7526732) Javier Cortés (426507) Mercè Martí (11711960) |
| author2_role | author author author author author author author author |
| author_facet | Andrea Aran (11711939) Gonzalo Lázaro (16752705) Vicente Marco (6068645) Vicente Peg (466433) Maitane Faus (22686683) Laia Garrigós (11644057) José Pérez-García (7526732) Javier Cortés (426507) Mercè Martí (11711960) |
| author_role | author |
| dc.creator.none.fl_str_mv | Andrea Aran (11711939) Gonzalo Lázaro (16752705) Vicente Marco (6068645) Vicente Peg (466433) Maitane Faus (22686683) Laia Garrigós (11644057) José Pérez-García (7526732) Javier Cortés (426507) Mercè Martí (11711960) |
| dc.date.none.fl_str_mv | 2025-11-26T05:15:41Z |
| dc.identifier.none.fl_str_mv | 10.3389/fimmu.2025.1699262.s001 |
| dc.relation.none.fl_str_mv | https://figshare.com/articles/dataset/Supplementary_file_1_Minimally_expanded_breast_cancer_tumor-infiltrating-lymphocytes_provide_guidance_for_therapeutic_selection_docx/30717896 |
| dc.rights.none.fl_str_mv | CC BY 4.0 info:eu-repo/semantics/openAccess |
| dc.subject.none.fl_str_mv | Genetic Immunology breast cancer TIL tumor-infiltrating lymphocytes immunotherapy TCR repertoire |
| dc.title.none.fl_str_mv | Supplementary file 1_Minimally expanded breast cancer tumor-infiltrating-lymphocytes provide guidance for therapeutic selection.docx |
| dc.type.none.fl_str_mv | Dataset info:eu-repo/semantics/publishedVersion dataset |
| description | Introduction<p>The analysis of tumor-infiltrating lymphocytes often requires techniques that expand their numbers, potentially introducing bias. To address this, we performed a detailed analysis of minimally cultured TILs to evaluate whether this approach better preserves their characteristics.</p>Methods<p>The TIL culture method was based solely on tumor tissue with low IL-2 supplementation to minimize artificial alterations. The validity of this approach was confirmed by the correlation between CD3+ T cell percentages in cultures and infiltration patterns observed by immunohistochemistry. Immunophenotyping, cytokine release, and TCR repertoire analysis were used to characterize CD4+ and CD8+ T cell subsets and their molecular features during minimal expansions.</p>Results<p>High TIL infiltration areas did not consistently correspond to an increased presence of any T cell subset; both CD4+ and CD8+ T cells frequently coexisted in these regions. In contrast, low TIL infiltration sections often displayed a higher proportion of CD4+ T cells. An inverse correlation between CD4+ T cell percentages and cytotoxic molecules was observed, indicating reduced cytotoxic activity in low-TIL sections with abundant CD4+ T cells. TCR repertoire analysis revealed differences between T cell subsets: CD4+ T cells were associated with longer TRA CDR3 nt and shorter TRB N(D)N nt lengths, along with lower diversity, while CD8+ T cells did not exhibit significant correlation with any TCR feature.</p>Discussion<p>This study highlights the distinct biological features of CD4⁺ and CD8⁺ TIL populations within the tumor microenvironment that can be preserved using a minimally expanded TIL approach. The observed associations between IHC patterns, T cell subset composition, cytotoxic potential, and TCR repertoire diversity help identify which biopsy regions yield TILs with greater therapeutic potential, thus providing guidance for TIL selection in immunotherapy.</p> |
| eu_rights_str_mv | openAccess |
| id | Manara_65092fa21872e6bf42868867e686d95a |
| identifier_str_mv | 10.3389/fimmu.2025.1699262.s001 |
| network_acronym_str | Manara |
| network_name_str | ManaraRepo |
| oai_identifier_str | oai:figshare.com:article/30717896 |
| publishDate | 2025 |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| rights_invalid_str_mv | CC BY 4.0 |
| spelling | Supplementary file 1_Minimally expanded breast cancer tumor-infiltrating-lymphocytes provide guidance for therapeutic selection.docxAndrea Aran (11711939)Gonzalo Lázaro (16752705)Vicente Marco (6068645)Vicente Peg (466433)Maitane Faus (22686683)Laia Garrigós (11644057)José Pérez-García (7526732)Javier Cortés (426507)Mercè Martí (11711960)Genetic Immunologybreast cancerTILtumor-infiltrating lymphocytesimmunotherapyTCR repertoireIntroduction<p>The analysis of tumor-infiltrating lymphocytes often requires techniques that expand their numbers, potentially introducing bias. To address this, we performed a detailed analysis of minimally cultured TILs to evaluate whether this approach better preserves their characteristics.</p>Methods<p>The TIL culture method was based solely on tumor tissue with low IL-2 supplementation to minimize artificial alterations. The validity of this approach was confirmed by the correlation between CD3+ T cell percentages in cultures and infiltration patterns observed by immunohistochemistry. Immunophenotyping, cytokine release, and TCR repertoire analysis were used to characterize CD4+ and CD8+ T cell subsets and their molecular features during minimal expansions.</p>Results<p>High TIL infiltration areas did not consistently correspond to an increased presence of any T cell subset; both CD4+ and CD8+ T cells frequently coexisted in these regions. In contrast, low TIL infiltration sections often displayed a higher proportion of CD4+ T cells. An inverse correlation between CD4+ T cell percentages and cytotoxic molecules was observed, indicating reduced cytotoxic activity in low-TIL sections with abundant CD4+ T cells. TCR repertoire analysis revealed differences between T cell subsets: CD4+ T cells were associated with longer TRA CDR3 nt and shorter TRB N(D)N nt lengths, along with lower diversity, while CD8+ T cells did not exhibit significant correlation with any TCR feature.</p>Discussion<p>This study highlights the distinct biological features of CD4⁺ and CD8⁺ TIL populations within the tumor microenvironment that can be preserved using a minimally expanded TIL approach. The observed associations between IHC patterns, T cell subset composition, cytotoxic potential, and TCR repertoire diversity help identify which biopsy regions yield TILs with greater therapeutic potential, thus providing guidance for TIL selection in immunotherapy.</p>2025-11-26T05:15:41ZDatasetinfo:eu-repo/semantics/publishedVersiondataset10.3389/fimmu.2025.1699262.s001https://figshare.com/articles/dataset/Supplementary_file_1_Minimally_expanded_breast_cancer_tumor-infiltrating-lymphocytes_provide_guidance_for_therapeutic_selection_docx/30717896CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/307178962025-11-26T05:15:41Z |
| spellingShingle | Supplementary file 1_Minimally expanded breast cancer tumor-infiltrating-lymphocytes provide guidance for therapeutic selection.docx Andrea Aran (11711939) Genetic Immunology breast cancer TIL tumor-infiltrating lymphocytes immunotherapy TCR repertoire |
| status_str | publishedVersion |
| title | Supplementary file 1_Minimally expanded breast cancer tumor-infiltrating-lymphocytes provide guidance for therapeutic selection.docx |
| title_full | Supplementary file 1_Minimally expanded breast cancer tumor-infiltrating-lymphocytes provide guidance for therapeutic selection.docx |
| title_fullStr | Supplementary file 1_Minimally expanded breast cancer tumor-infiltrating-lymphocytes provide guidance for therapeutic selection.docx |
| title_full_unstemmed | Supplementary file 1_Minimally expanded breast cancer tumor-infiltrating-lymphocytes provide guidance for therapeutic selection.docx |
| title_short | Supplementary file 1_Minimally expanded breast cancer tumor-infiltrating-lymphocytes provide guidance for therapeutic selection.docx |
| title_sort | Supplementary file 1_Minimally expanded breast cancer tumor-infiltrating-lymphocytes provide guidance for therapeutic selection.docx |
| topic | Genetic Immunology breast cancer TIL tumor-infiltrating lymphocytes immunotherapy TCR repertoire |