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Clin Cancer Res. 2016 Feb 1;22(3):704-13. doi: 10.1158/1078-0432.CCR-15-1543. Epub 2015 Sep 25.

Evaluation of PD-L1 Expression and Associated Tumor-Infiltrating Lymphocytes in Laryngeal Squamous Cell Carcinoma.

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Department of Pathology, Yale University School of Medicine, New Haven, Connecticut.
Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Athens, Athens, Greece.
Department of Pathology, Aristotle University of Thessaloniki School of Medicine, Thessaloniki, Greece. Laboratory of Molecular Oncology, Hellenic Foundation for Cancer Research, Thessaloniki, Greece.
Department of Surgery (Otolaryngology), Yale University School of Medicine, New Haven, Connecticut.
Laboratory of Molecular Oncology, Hellenic Foundation for Cancer Research, Thessaloniki, Greece.
Department of Cranio-, Maxillofacial and Oral Surgery, Medical University of Vienna, Vienna, Austria.
First Ear, Nose and Throat Clinics, Athens General Hospital "Hippokration," University of Athens, Athens, Greece.
Second Department of Internal Medicine, Section of Medical Oncology, "Attikon" University Hospital, University of Athens, Athens, Greece.



Programmed death-ligand 1 (PD-L1; also known as CD274 or B7-H1) expression represents a mechanism of immune escape for cancer. Our purpose was to characterize tumor PD-L1 expression and associated T-cell infiltration in primary laryngeal squamous cell carcinomas (SCC).


A well-annotated cohort of 260 operable primary laryngeal SCCs [formalin-fixed paraffin-embedded (FFPE) specimens] was morphologically characterized for stromal tumor-infiltrating lymphocytes (TIL), on hematoxylin/eosin-stained whole sections and for PD-L1 mRNA expression by qRT-PCR in FFPE specimens. For PD-L1 protein expression, automated quantitative protein analysis (AQUA) was applied on tissue microarrays consisting of two cores from these tumors. In addition, PD-L1 mRNA expression in fresh-frozen tumors and normal adjacent tissue specimens was assessed in a second independent cohort of 89 patients with primary laryngeal SCC.


PD-L1 mRNA levels were upregulated in tumors compared with surrounding normal tissue (P = 0.009). TILs density correlated with tumor PD-L1 AQUA levels (P = 0.021). Both high TILs density and high PD-L1 AQUA levels were significantly associated with superior disease-free survival (DFS; TILs: P = 0.009 and PD-L1: P = 0.044) and overall survival (OS; TILs: P = 0.015 and PD-L1: P = 0.059) of the patients and retained significance in multivariate analysis.


Increased TILs density and PD-L1 levels are associated with better outcome in laryngeal squamous cell cancer. Assessment of TILs and PD-L1 expression could be useful to predict response to immune checkpoint inhibitors.

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