Department of Otorhinolaryngology - Head & Neck Surgery, Medical University of Innsbruck, Anichstrasse 35, Innsbruck, 6020, Austria.
University Clinics Innsbruck, Tirol Kliniken, Anichstrasse 35, Innsbruck, 6020, Austria.
BMC Cancer. 2023 Nov 27;23(1):1154. doi: 10.1186/s12885-023-11440-x.
Epithelial, connective tissue and immune cells contribute in various ways to the pathophysiology of HPV positive (HPV+) and HPV negative (HPV-) oropharyngeal squamous cell carcinoma (OPSCC). We aimed to investigate the abundance of these cell lineages and their coexpression patterns in patients with HPV + and HPV- OPSCC.
We used a 4-channel immunofluorescence-microscopy technique for the simultaneous detection of three direct-conjugated antibodies (pancytokeratin, vimentin and CD45/CD18) and DAPI (4',6-Diamidin-2-phenylindole) in formalin fixed paraffin-embedded tissue samples (FFPE) of patients with HPV + and HPV- OPSCC, and of control patients. Image acquisition and analysis were performed with TissueFAXS and StrataQuest (TissueGnostics, Vienna, Austria), respectively, in tumor cell clusters/stroma in OPSCC specimens and epithelial layer/lamina propria in control specimens. Cell populations were created based on antibodies' coexpression patterns. Isotype and positive controls were examined for plausibility.
The proportion of cells of epithelial differentiation in tumor cell clusters was higher in HPV + OPSCC (55%) than in HPV- OPSCC samples (44%). The proportion of connective tissue cells in tumor cell cluster was lower in HPV + OPSCC patients (18%) than in HPV- OPSCC patients (26%). The proportion of immune cells in tumor cell clusters was higher in HPV + OPSCC patients (25%) than in HPV- OPSCC patients (18%). The percentage of anaplastic, potentially de-differentiated cells, was 2% in control patients, and it was higher in HPV- OPSCC (21%) than in HPV + OPSCC samples (6%).
This study provided the first quantitative data for the abundance of cells of epithelial, connective tissue and immune differentiation, in patients with OPSCC and control patients. The abundance of these different crucial cell populations was consistently originating from the same tissue sample. De-differentiation of tumor cells was higher in HPV- OPSCC than in HPV + OPSCC. In tumor cells clusters, the antitumoral host immune response was higher in HPV + OPSCC than in HPV- OPSCC, whereas the fibroblast response was higher in HPV- OPSCC than in HPV + OPSCC. This study contributed to the understanding of histopathologic differences between HPV + OPSCC and HPV- OPSCC patients.
上皮细胞、结缔组织细胞和免疫细胞以不同的方式参与 HPV 阳性（HPV+）和 HPV 阴性（HPV-）口咽鳞状细胞癌（OPSCC）的病理生理学。我们旨在研究这些细胞谱系在 HPV+和 HPV-OPSCC 患者中的丰度及其共表达模式。
我们使用 4 通道免疫荧光显微镜技术，同时检测 HPV+和 HPV-OPSCC 患者以及对照患者福尔马林固定石蜡包埋（FFPE）组织样本中的三种直接缀合抗体（细胞角蛋白、波形蛋白和 CD45/CD18）和 DAPI（4'，6-二脒基-2-苯基吲哚）。图像采集和分析分别使用 TissueFAXS 和 StrataQuest（TissueGnostics，维也纳，奥地利）进行，在 OPSCC 标本中的肿瘤细胞簇/基质中和对照标本中的上皮层/固有层中进行。基于抗体的共表达模式创建细胞群体。检查同型和阳性对照以确保其合理性。
HPV+OPSCC 肿瘤细胞簇中上皮分化细胞的比例（55%）高于 HPV-OPSCC 样本（44%）。HPV+OPSCC 患者肿瘤细胞簇中的结缔组织细胞比例（18%）低于 HPV-OPSCC 患者（26%）。HPV+OPSCC 患者肿瘤细胞簇中的免疫细胞比例（25%）高于 HPV-OPSCC 患者（18%）。对照患者中潜在去分化的间变细胞的百分比为 2%，而 HPV-OPSCC 患者（21%）高于 HPV+OPSCC 样本（6%）。
本研究首次提供了 OPSCC 患者和对照患者上皮、结缔组织和免疫分化细胞丰度的定量数据。这些不同关键细胞群体的丰度均源自同一组织样本。HPV-OPSCC 中肿瘤细胞的去分化程度高于 HPV+OPSCC。在肿瘤细胞簇中，HPV+OPSCC 中的抗肿瘤宿主免疫反应高于 HPV-OPSCC，而 HPV-OPSCC 中的成纤维细胞反应高于 HPV+OPSCC。本研究有助于理解 HPV+OPSCC 和 HPV-OPSCC 患者之间的组织病理学差异。
