Pokrývková Barbora, Šmahelová Jana, Dalewská Natálie, Grega Marek, Vencálek Ondřej, Šmahel Michal, Nunvář Jaroslav, Klozar Jan, Tachezy Ruth
Department of Genetics and Microbiology, Faculty of Science, Charles University, BIOCEV, 252 50 Vestec, Czech Republic.
Department of Pathology and Molecular Medicine, 2nd Faculty of Medicine, Charles University, 150 06 Prague, Czech Republic.
Diagnostics (Basel). 2021 Mar 31;11(4):628. doi: 10.3390/diagnostics11040628.
Head and neck squamous cell carcinomas (HNSCC) can be induced by smoking or alcohol consumption, but a growing part of cases relate to a persistent high-risk papillomavirus (HPV) infection. Viral etiology has a beneficial impact on the prognosis, which may be explained by a specific immune response. Tumor associated macrophages (TAMs) represent the main immune population of the tumor microenvironment with a controversial influence on the prognosis. In this study, the level, phenotype, and spatial distribution of TAMs were evaluated, and the expression of TAM-associated markers was compared in HPV positive (HPV+) and HPV negative (HPV-) tumors. Seventy-three formalin and embedded in paraffin (FFPE) tumor specimens were examined using multispectral immunohistochemistry for the detection of TAM subpopulations in the tumor parenchyma and stroma. Moreover, the mRNA expression of TAM markers was evaluated using RT-qPCR. Results were compared with respect to tumor etiology, and the prognostic significance was evaluated. In HPV- tumors, we observed more pro-tumorigenic M2 in the stroma and a non-macrophage arginase 1 (ARG1)-expressing population in both compartments. Moreover, higher mRNA expression of M2 markers-cluster of differentiation 163 (CD163), ARG1, and prostaglandin-endoperoxide synthase 2 (PTGS2)-was detected in HPV- patients, and of M1 marker nitric oxide synthase 2 (NOS2) in HPV+ group. The expression of ARG1 mRNA was revealed as a negative prognostic factor for overall survival of HNSCC patients.
头颈部鳞状细胞癌（HNSCC）可由吸烟或饮酒诱发，但越来越多的病例与持续性高危人乳头瘤病毒（HPV）感染有关。病毒病因对预后有有益影响，这可能由特定的免疫反应来解释。肿瘤相关巨噬细胞（TAM）是肿瘤微环境中的主要免疫细胞群体，对预后的影响存在争议。在本研究中，评估了TAM的水平、表型和空间分布，并比较了HPV阳性（HPV+）和HPV阴性（HPV-）肿瘤中TAM相关标志物的表达。使用多光谱免疫组织化学检查了73个福尔马林固定石蜡包埋（FFPE）肿瘤标本，以检测肿瘤实质和间质中的TAM亚群。此外，使用RT-qPCR评估了TAM标志物的mRNA表达。将结果与肿瘤病因进行比较，并评估其预后意义。在HPV-肿瘤中，我们在间质中观察到更多促肿瘤的M2型巨噬细胞，且在两个区域均观察到一个不表达巨噬细胞精氨酸酶1（ARG1）的细胞群体。此外，在HPV-患者中检测到M2标志物——分化簇163（CD163）、ARG1和前列腺素内过氧化物合酶2（PTGS2）的mRNA表达较高，而在HPV+组中检测到M1标志物一氧化氮合酶2（NOS2）的mRNA表达较高。ARG1 mRNA的表达被揭示为HNSCC患者总生存的负性预后因素。
