Primary head and neck cancer cell cultures are susceptible to proliferation of Epstein-Barr virus infected lymphocytes

BMC Cancer

Research

Laura Martínez-Ruiz¹  Javier Florido¹  Germaine Escames¹  Carsten Hain²  Pascal Schmidt²  Jörn Kalinowski²  Michael Wehmeier³  Holger Sudhoff⁴  Felix Oppel⁴  Peter Goon⁴  Matthias Schürmann⁴  Senyao Shao⁴  Lars Uwe Scholtz⁴  Sarah Gendreizig⁴  Menghang Wang⁵  Leonie Hose⁶  Frank Brasch⁷  Almut Mentz⁷ 

[1] Biomedical Research Center, Health Sciences Technology Park, University of Granada, 18016, Granada, Spain;Department of Physiology, Faculty of Medicine, University of Granada, 18016, Granada, Spain;CIBERFES, Ibs. Granada, San Cecilio University Hospital, 18016, Granada, Spain;Center for Biotechnology (CeBiTec), Universität Bielefeld, Bielefeld, Germany;Department of Laboratory Medicine, Klinikum Bielefeld, Teutoburger Str. 50, 33604, Bielefeld, Germany;Department of Otolaryngology, Head and Neck Surgery, Campus Klinikum Bielefeld Mitte, University Hospital OWL of Bielefeld University, Klinikum Bielefeld, Teutoburger Str. 50, 33604, Bielefeld, Germany;Department of Otolaryngology, Head and Neck Surgery, Campus Klinikum Bielefeld Mitte, University Hospital OWL of Bielefeld University, Klinikum Bielefeld, Teutoburger Str. 50, 33604, Bielefeld, Germany;Department of Otolaryngology Head and Neck Surgery, Peking University International Hospital, Peking University, 102206, Beijing, China;Department of Otolaryngology, Head and Neck Surgery, Campus Klinikum Bielefeld Mitte, University Hospital OWL of Bielefeld University, Klinikum Bielefeld, Teutoburger Str. 50, 33604, Bielefeld, Germany;Department of Pathology, Klinikum Bielefeld, Teutoburger Str. 50, 33604, Bielefeld, Germany;Department of Pathology, Klinikum Bielefeld, Teutoburger Str. 50, 33604, Bielefeld, Germany;

关键词: Head and neck cancer;    Lymphocyte;    Epstein-barr virus;    Human papilloma virus;    Primary cell culture model;   

DOI  :  10.1186/s12885-022-10481-y

 received in 2021-04-01, accepted in 2022-12-23,  发布年份 2022

来源: Springer

【 摘 要 】

BackgroundNew concepts for a more effective anti-cancer therapy are urgently needed. Experimental flaws represent a major counter player of this development and lead to inaccurate and unreproducible data as well as unsuccessful translation of research approaches into clinics. In a previous study we have created epithelial cell cultures from head and neck squamous cell carcinoma (HNSCC) tissue.MethodsWe characterize primary cell populations isolated from human papillomavirus positive HNSCC tissue for their marker expression by RT-qPCR, flow cytometry, and immunofluorescence staining. Their sensitivity to MDM2-inhibition was measured using cell viability assays.ResultsPrimary HNSCC cell cultures showed the delayed formation of spheroids at higher passages. These spheroids mimicked the morphology and growth characteristics of other established HNSCC spheroid models. However, expression of epithelial and mesenchymal markers could not be detected in these cells despite the presence of the HNSCC stem cell marker aldehyde dehydrogenase 1 family member A1. Instead, strong expression of B- and T-lymphocytes markers was observed. Flow cytometry analysis revealed a heterogeneous mixture of CD3 + /CD25 + T-lymphocytes and CD19 + B-lymphocytes at a ratio of 4:1 at passage 5 and transformed lymphocytes at late passages (≥ passage 12) with CD45 + CD19 + CD20 + , of which around 10 to 20% were CD3 + CD25 + CD56 + . Interestingly, the whole population was FOXP3-positive indicative of regulatory B-cells (Bregs). Expression of transcripts specific for the Epstein-Barr-virus (EBV) was detected to increase in these spheroid cells along late passages, and this population was vulnerable to MDM2 inhibition. HPV + HNSCC cells but not EBV + lymphocytes were detected to engraft into immunodeficient mice.ConclusionsIn this study we present a primary cell culture of EBV-infected tumor-infiltrating B-lymphocytes, which could be used to study the role of these cells in tumor biology in future research projects. Moreover, by describing the detailed characteristics of these cells, we aim to caution other researchers in the HNSCC field to test for EBV-infected lymphocyte contaminations in primary cell cultures ahead of further experiments. Especially researchers who are interested in TIL-based adopted immunotherapy should exclude these cells in their primary tumor models, e.g. by MDM2-inhibitor treatment. BI-12-derived xenograft tumors represent a suitable model for in vivo targeting studies.

【 授权许可】

CC BY   
© The Author(s) 2023

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