The prognostic significance of peritumoral tertiary lymphoid structures in breast cancer
Tumors and their surrounding area represent spatially organized “ecosystems”, where tumor cells and the immune contextures of the different compartments are in a dynamic interplay, with potential clinical impact. Here, we aimed to investigate the prognostic significance of peritumoral tertiary lymphoid structures (TLS) either alone or jointly with the intratumoral densities and spatial distribution of CD8 + and CD163 + cells in breast cancer (BCa) patients. TLS were identified peritumorally, within the area distancing up to 5 mm from the infiltrative tumor border, counted and further characterized as adjacent or distal, in formalin-fixed, paraffin-embedded tumor tissue samples from a cohort of 167 patients, with histologically confirmed invasive ductal BCa. TLS and tumor-infiltrating immune cells were determined by H&E and immunohistochemistry. Clinical follow-up was available for 112 of these patients. Patients with peritumoral TLS exhibited worse disease-free survival (DFS) and overall survival (OS) as compared to patients lacking TLS. Moreover, the density of peritumoral TLS was found to be crucial for prognosis, since patients with abundant TLS exhibited the worst DFS and OS. By combining the density of adjacent TLS (aTLS) with our recently published intratumoral signatures based on the differential distribution of CD8 + and CD163 + in the tumor center and invasive margin, we created two improved immune signatures with superior prognostic strength and higher patient population coverage. Our observations strengthen the notion for the fundamental role of the dynamic interplay between the immune cells within the tumor microenvironment (center/invasive margin) and the tumor surrounding area (peritumoral TLS) on the clinical outcome of BCa patients.
KeywordsBreast cancer Tertiary lymphoid structure Tumor infiltration Immune contexture Prognostic signatures
American Joint Committee on Cancer
Adjacent tertiary lymphoid structure
Distant tertiary lymphoid structure
Favorable combined immune signature
Formalin fixed, paraffin embedded
High endothelial venule
Reinforced favorable combined immune signature
Reinforced unfavorable combined immune signature
Secondary lymphoid organ
Tertiary lymphoid structure
T regulatory cell
Unfavorable combined immune signature
MS and SPF equally contributed to study design, data collection, analysis and interpretation of results, and the writing the manuscript; CKV contributed to the study design, data collection, and interpretation of results; NNS contributed to data collection and analysis; AA and NA were responsible for the collection of patient material and clinical follow-up; DV contributed to data analysis and writing of the manuscript; SAP and CNB supervised the study, contributed to experimental design, data analysis, and wrote the manuscript. All authors read and approved the final version of the manuscript.
This study was supported by Grant GER_1968 (acronym ISPEBREAST) to Constantin N. Baxevanis from the Greek–German bilateral cooperation for research and innovation, funded by the General Secretariat for Research and Technology (GSRT) of the Ministry of Education, Research and Religious Affairs of the Hellenic Republic and the German Federal Ministry for Education and Research (BMBF), and by a donation to Sonia Perez from the Haegeman-Goossens family, Netherlands. We would like to acknowledge the Haegeman-Goossens family for their support to our research.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
Ethical approval and ethical standards
The study was approved by the Institutional Review Board of Saint Savas Cancer Hospital, Study code: IRB-ID 6079/448/10-6-13. Date of approval: 10/06/2013; retrospective study (2000–2010); first patient prospectively enrolled: 14/2/2014.
Retrospectively analyzed (2000–2010) and prospectively enrolled patients (2014–2015) who signed the same informed consent. Individual participants provided written informed consent for the use of their specimens. In cases where retrospectively analyzed patients could not visit the hospital to provide signed informed consent, they were informed orally via telephone calls by their oncologists. Clinical data were obtained from all patients anonymously. Patients were assured that confidentiality of their records will be protected according to the Greek regulations and laws.
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