CD4+ and CD8+ T cells in sentinel nodes exhibit distinct pattern of PD‐1, CD69, and HLA‐DR expression compared to tumor tissue in oral squamous cell carcinoma

• Published in: Cancer science 2021-03, Vol.112 (3), p.1048-1059
• Main Authors: Piersiala, Krzysztof, Farrajota Neves da Silva, Pedro, Hjalmarsson, Eric, Kolev, Aeneas, Kågedal, Åsa, Starkhammar, Magnus, Elliot, Alexandra, Marklund, Linda, Margolin, Gregori, Munck‐Wikland, Eva, Kumlien Georén, Susanna, Cardell, Lars‐Olaf
• Format: Article
• Language: English
• Subjects: Adult; Aged; Aged, 80 and over; Antigens; Antigens, CD - analysis; Antigens, CD - metabolism; Antigens, Differentiation, T-Lymphocyte - analysis; Antigens, Differentiation, T-Lymphocyte - metabolism; Apoptosis; Biomarkers; Cancer; CD3 antigen; CD4 antigen; CD4-Positive T-Lymphocytes - immunology; CD4-Positive T-Lymphocytes - metabolism; CD69 antigen; CD8 antigen; CD8-Positive T-Lymphocytes - immunology; CD8-Positive T-Lymphocytes - metabolism; Cell death; Cytotoxicity; Dimensional analysis; Ethics; Female; Flow Cytometry; head and neck cancer; Histocompatibility antigen HLA; HLA-DR Antigens - analysis; HLA-DR Antigens - metabolism; Humans; Immunology; Immunotherapy; Lectins, C-Type - analysis; Lectins, C-Type - metabolism; Leukocytes (mononuclear); Lymph nodes; Lymphatic system; Lymphocytes; Lymphocytes T; Lymphocytes, Tumor-Infiltrating - immunology; Lymphocytes, Tumor-Infiltrating - metabolism; Male; Medical prognosis; Medicin och hälsovetenskap; Metastasis; metastatic head and neck squamous cell carcinoma; Middle Aged; Mouth Neoplasms - immunology; Mouth Neoplasms - pathology; Oral squamous cell carcinoma; Original; Patients; PD‐1; Peripheral blood mononuclear cells; Programmed Cell Death 1 Receptor - analysis; Programmed Cell Death 1 Receptor - metabolism; R&D; Research & development; Sentinel Lymph Node - cytology; Sentinel Lymph Node - immunology; Sentinel Lymph Node - pathology; sentinel node; Software; Squamous cell carcinoma; Squamous Cell Carcinoma of Head and Neck - immunology; Squamous Cell Carcinoma of Head and Neck - pathology; Survival analysis; tumor‐draining lymph nodes
• ISSN: 1347-9032; 1349-7006; 1349-7006
• DOI: 10.1111/cas.14816

Anticancer immunotherapies have revolutionized cancer management, yet the effect of systemic anti‐programmed cell death protein 1 (PD‐1) treatment is predominantly studied in tumor‐infiltrating lymphocytes (TILs). Its impact on PD‐1 expressing cells in tumor‐draining lymph nodes (TDLNs) is not well understood and yet to be explored. Thus, further research aiming for better understanding of the PD‐1 pathway not only in tumor tissue but also in TDLNs is warranted. In this study, we investigated the expression of PD‐1, CD69, and HLA‐DR on CD4+ and CD8+ T cells by flow cytometry analysis of peripheral blood mononuclear cells (PBMCs), TDLNs, and tumor samples from patients with oral squamous cell carcinoma (OSCC). Our data showed that both helper and cytotoxic T lymphocytes in OSCC tissue were highly activated and expressed high level of PD‐1 (over 70% positivity). Lymphocytes in TDLNs and peripheral blood expressed significantly lower levels of PD‐1 and other activation markers compared to TILs. Moreover, we demonstrated that a significant fraction of PD‐1 negative TILs expressed high levels of human leukocyte antigen – DR isotype and CD69. In contrast, PD‐1 negative cells in TDLNs and PBMCs scarcely expressed the aforementioned activation markers. Furthermore, we proved that patients with a high percentage of CD3+ PD‐1+ cells in tumor‐draining lymph nodes had significantly lower disease‐free and overall survival rates (log‐rank test P = .0272 and P = .0276, respectively). Taken together, we proved that flow cytometry of lymph nodes in OSCC is feasible and may be used to investigate whether PD‐1 levels in TDLNs correspond with survival and potentially with response to anti‐PD‐1 therapy. Such knowledge may ultimately help guide anti‐PD‐1 treatment. Flow cytometry is a powerful tool that can be used in head and neck cancers to investigate immunological milieu of tumor‐draining lymph nodes. In this project, we provide a characterization of T cells within head and neck cancer tumors, tumor‐draining lymph nodes, and blood.