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This study investigates the relationship between tumour-infiltrating lymphocytes and the vascular compartment in basal-type triple negative breast cancer. Using multiplex fluorescent immunohistochemistry and quantitative digital analysis in 10 basal tumours, CD3+ T cells were assessed relative to blood vessels and endothelial activation markers (VCAM-1, ICAM-1, PD-L1). In silico validation used TCGA BRCARNA-seq data (N=808), showing elevated lymphocyte numbers associated with endothelial activation.",{"@graph":69,"@context":122},[70,84,105],{"@type":71,"itemListElement":72},"BreadcrumbList",[73,77,79,82],{"item":74,"name":75,"@type":76,"position":8},"https://docshare.wps.com","Home","ListItem",{"item":78,"name":9,"@type":76,"position":14},"https://docshare.wps.com/document/",{"item":80,"name":35,"@type":76,"position":81},"https://docshare.wps.com/document/healthcare/",3,{"item":83,"name":65,"@type":76,"position":19},"https://docshare.wps.com/document/preliminary-characterisation-of-the-spatial-immune-and-vascular-environment-in-triple-negative-basal-breast-carcinomas-using-multiplex-fluorescent-immunohistochemistry-research-article/370163/",{"url":83,"name":65,"@type":85,"image":86,"author":91,"headline":65,"publisher":94,"fileFormat":97,"inLanguage":63,"description":67,"dateModified":98,"datePublished":99,"encodingFormat":97,"isAccessibleForFree":100,"interactionStatistic":101},"DigitalDocument",{"url":87,"@type":88,"width":89,"height":90},"https://docshare.wps.com/thumbnails/preliminary-characterisation-of-the-spatial-immune-and-vascular-environment-in-triple-negative-basal-breast-carcinomas-using-multiplex-fluorescent-immunohistochemistry-research-article/370163.png","ImageObject",300,407,{"name":92,"@type":93},"Logic","Person",{"url":74,"name":95,"@type":96},"DocShare","Organization","application/pdf","2026-10-27","2026-09-23",true,{"@type":102,"interactionType":103,"userInteractionCount":52},"InteractionCounter",{"@type":104},"ViewAction",{"@type":106,"mainEntity":107},"FAQPage",[108,114,118],{"name":109,"@type":110,"acceptedAnswer":111},"What does the study examine in triple negative basal breast carcinomas?","Question",{"text":112,"@type":113},"It examines how tumour-infiltrating lymphocytes relate to blood vessels and endothelial activation, aiming to clarify mechanisms of immune cell trafficking in basal-type TNBC.","Answer",{"name":115,"@type":110,"acceptedAnswer":116},"How were immune and vascular features measured?",{"text":117,"@type":113},"Spatial immune features were characterized in 10 basal breast cancers using multiplex fluorescent immunohistochemistry and quantitative digital analysis of CD3+ T cells, with endothelial activation assessed via VCAM-1, ICAM-1 and PD-L1.",{"name":119,"@type":110,"acceptedAnswer":120},"What did the study find about CD3+ T cell densities and vessel activation?",{"text":121,"@type":113},"CD3+ T cells had higher densities in the stromal compartment than the neoplastic compartment. ICAM-1-activated blood vessels correlated with higher CD3+ T cell densities only within 30 microns of vessels.","https://schema.org",{"og:url":83,"og:type":124,"og:title":65,"og:site_name":95,"og:description":67},"article",{"robots":126,"canonical":83},"index,follow",{"doc_id":128,"site_id":62},370163,1790404576,{"code":4,"msg":5,"data":131},{"doc_id":128,"user_id":132,"nickname":92,"user_avatar":133,"doc_module":4,"category_id":34,"category_name":35,"doc_title":65,"doc_description":67,"doc_content":134,"file_id":135,"file_url":136,"file_type":137,"file_size":138,"view_count":52,"is_deleted":4,"is_public":8,"is_downloadable":8,"audit_status":8,"page_count":139,"language":140,"language_code":63,"site_id":62,"html_lang":63,"table_of_contents":141,"faqs":142,"seo_title":143,"seo_description":67,"update_tm":144,"read_time":145},1099513958762,"https://ap-avatar.wpscdn.com/avatar/1000023916a998db790?x-image-process=image/resize,m_fixed,w_180,h_180&k=1784791008015729253","PLOS ONE  \nOPEN ACCESS  \nCitation: Takano EA, Jana MK, Lara Gonzalez LE, Pang J-MB, Salgado R, Loi S, et al. (2025) Preliminary characterisation of the spatial immune and vascular environment in triple negative basal breast carcinomas using multiplex fluorescent immunohistochemistry. PLoS ONE 20(1):  \ne0317331 . [https://doi.org/10.1371/journal](https://doi.org/10.1371/journal). pone.0317331  \nEditor: Lu Zhang, Fudan University, CHINA  \nReceived: June 2, 2024  \nAccepted: December 27, 2024  \nPublished: January 10, 2025  \nPeer Review History: PLOS recognizes the benefits of transparency in the peer review process; therefore, we enable the publication of all of the content of peer review and author responses alongside final, published articles. The editorial history of this article is available here:  \n[https://doi.org/10.1371/journal.pone.0317331](https://doi.org/10.1371/journal.pone.0317331)  \n[Copyright:](Copyright:) © [2025 Takano et al](2025 Takano et al). This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.  \nData Availability Statement: All relevant data are within the manuscript and its Supporting Information files.  \nRESEARCH ARTICLE  \nPreliminary characterisation of the spatial immune and vascular environment in triple negative basal breast carcinomas using multiplex fluorescent immunohistochemistry  \nElena A. Takano1 *, Metta K. Jana2, Luis E. Lara Gonzalez3,4, Jia-Min B. Pang1, Roberto Salgado3,5, Sherene Loi3,4, Stephen B. Fox1,4 *  \n1 Department of Pathology, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia, 2 Centre for Advanced Histology and Microscopy, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia,  \n3 Cancer Research Division, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia, 4 The Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia,  \n5 GZA-ZNA-Hospitals, Antwerp, Belgium  \n* [elena.takano@petermac.org](elena.takano@petermac.org) (EAT); [stephen.fox@petermac.org](stephen.fox@petermac.org) (SBF)  \nAbstract  \nTriple negative breast cancers often contain higher numbers of tumour-infiltrating lymphocytes compared with other breast cancer subtypes, with their number correlating with prolonged survival. Since little is known about tumour-infiltrating lymphocyte trafficking in triple negative breast cancers, we investigated the relationship between tumour-infiltrating lymphocytes and the vascular compartment to better understand the immune tumour microenvironment in this aggressive cancer type. We aimed to identify mechanisms and signaling pathways responsible for immune cell trafficking in triple negative breast cancers, specifically of basal type, that could potentially be manipulated to change such tumours from immune “cold” to “hot” thereby increasing the likelihood of successful immunotherapy in this challenging patient population. We characterised the spatial immune environment in 10 basal breast cancers showing a range of tumour-infiltrating lymphocytes using multiplex fluorescent immunohistochemistry and quantitative digital analysis of CD3+ T cells. We examined their relationship to blood vessels and their activation status as defined by VCAM- 1, ICAM-1 and PD-L1 . Confirmation of the relationship between tumour-infiltrating lymphocytes and endothelial activation was performed through insilico analysis on TCGA BRCARNA-seq data (N = 808) . Significantly higher CD3+ T cell densities were observed in the stromal compartment compared with the neoplastic cell compartment (P = 0.003) . ICAM-1 activated blood vessels were spatially associated with higher CD3+ T cell densities only within 30 microns of blood vessels compared with more distal activated and non-activated blood vessels (P = 0.041) . In silico analysis confirmed higher numbers of tu","cbCaicb8na83Xrod","https://ap.wps.com/l/cbCaicb8na83Xrod","pdf",3020981,18,"English","# Abstract\n# Introduction\n# Research approach and findings\n## Spatial analysis of immune cells and vessels\n## Endothelial activation and PD-L1 regulation","[{\"question\":\"What does the study examine in triple negative basal breast carcinomas?\",\"answer\":\"It examines how tumour-infiltrating lymphocytes relate to blood vessels and endothelial activation, aiming to clarify mechanisms of immune cell trafficking in basal-type TNBC.\"},{\"question\":\"How were immune and vascular features measured?\",\"answer\":\"Spatial immune features were characterized in 10 basal breast cancers using multiplex fluorescent immunohistochemistry and quantitative digital analysis of CD3+ T cells, with endothelial activation assessed via VCAM-1, ICAM-1 and PD-L1.\"},{\"question\":\"What did the study find about CD3+ T cell densities and vessel activation?\",\"answer\":\"CD3+ T cells had higher densities in the stromal compartment than the neoplastic compartment. ICAM-1-activated blood vessels correlated with higher CD3+ T cell densities only within 30 microns of vessels.\"}]","Preliminary characterisation of the spatial immune and vascular environment in triple negative basal breast carcinomas using multiplex fluorescent immunohistochemistry - Research Article | PDF",1790187124,45]