Tumour-draining lymph nodes in head and neck cancer are characterized by accumulation of CTLA-4 and PD-1 expressing Treg cells

Krzysztof Piersiala; Pedro Farrajota Neves da Silva; Vilma Lagebro; Aeneas Kolev; Magnus Starkhammar; Alexandra Elliot; Linda Marklund; Eva Munck-Wikland; Gregori Margolin; Susanna Kumlien Georén; Lars-Olaf Cardell

doi:10.1016/j.tranon.2022.101469

Tumour-draining lymph nodes in head and neck cancer are characterized by accumulation of CTLA-4 and PD-1 expressing Treg cells

Transl Oncol. 2022 Sep:23:101469. doi: 10.1016/j.tranon.2022.101469. Epub 2022 Jun 14.

Affiliations

¹ Division of ENT Diseases, Department of Clinical Sciences, Intervention and Technology, Karolinska Institutet, Stockholm; Department of Otorhinolaryngology, Karolinska University Hospital, Stockholm, Sweden.
² Department of Pathology and Cytology, Karolinska University Hospital, Stockholm, Sweden.
³ Division of ENT Diseases, Department of Clinical Sciences, Intervention and Technology, Karolinska Institutet, Stockholm.
⁴ Division of ENT Diseases, Department of Clinical Sciences, Intervention and Technology, Karolinska Institutet, Stockholm; Department of Otorhinolaryngology, Karolinska University Hospital, Stockholm, Sweden; Medical unit Head Neck, Lung and skin Cancer, Karolinska University Hospital, Stockholm, Sweden.
⁵ Division of ENT Diseases, Department of Clinical Sciences, Intervention and Technology, Karolinska Institutet, Stockholm; Department of Otorhinolaryngology, Karolinska University Hospital, Stockholm, Sweden. Electronic address: lars-olaf.cardell@ki.se.

Abstract

Introduction: High Tregs infiltration within the tumour microenvironment (TME) of various cancers shows a positive correlation with poor prognosis. Despite the fact that tumour draining lymph nodes (TDLNs) are recognized as key organs playing a crucial role in response to immunotherapy and modulating anti-cancer immunity, the distribution of Tregs and their role in TDLNs remain uncertain thus far. The purpose of this project is to investigate the density of Tregs in TDLNs and non-TDLNs and their expression of immune checkpoint molecules - PD-1 and CTLA-4.

Methods: Samples including TDLNs, non-TDLNs and metastatic lymph nodes (LNs) from 23 patients with oral squamous cell carcinoma (OSCC) were analyzed by multicolour flow cytometry with a focus on Tregs population and expression of CTLA-4 and PD-1.

Results: TDLNs and metastatic LNs were characterized by a significantly higher infiltration of Tregs defined as CD4+FoxP3+CD25^highCD127^low cells and significantly higher expression of CTLA-4 and PD-1 on Tregs compared with non-TDLNs. Tregs in TDLNs and metastatic LNs co-expressed CTLA-4 and PD-1 abundantly. High expression of these immune check-point molecules correlated with positive N-stage but not with T-stage.

Conclusion: TDLNs and metastatic LNs are characterized by a high accumulation of Tregs expressing high levels of CTLA-4 and PD-1. High infiltration of Tregs can be a potential driver of an immunosuppressive milieu in TDLNs that can, in turn, favour cancer progression. High accumulation of Tregs expressing CTLA-4 and PD-1 in TDLNs is associated with lymph node involvement, but not with the size of the primary tumour.

Keywords: CTLA-4; HNSCC; Immunotherapy; PD-1; T regulatory cells.