Introduction: Human papillomavirus with a high oncogenic potential (HR-HPV) is responsible for more than a half of squamous cell carcinomas of the oropharynx. The HR-HPV-dependent cases of this tumour have a better prognosis compared to the HR-HPV-negative cases, despite the usually more advanced disease at the time of diagnosis. In addition to genetic and epigenetic factors, the causes of this more favourable course of the disease are also seen in the participation of the tumour microenvironment, including the patient’s immune system. Macrophages are one of the most important elements of the immunocompetent cells landscape that make up the tumour microenvironment. Traditionally, they are divided into 2 groups: inflammatory macrophages with the M1 phenotype and tumour-associated macrophages known as M2 phenotype macrophages. Objective: The aim of this study was to investigate the impact of the macrophage infiltrates intensity of the M1/M2 and M2 phenotype separately on the clinical outcome of patients with squamous cell carcinoma of the oropharynx (OPSCC), taking into account the HR-HPV status of tumours. Methods: The study involved 85 patients with OPSCC in which HR-HPV status in tumour tissue was determined using a double-check algorithm including the detection of viral DNA by RT-PCR method with subsequent confirmation of its biological activity by immunohistochemical demonstrating the P16INK4A protein overexpression. In each of the groups formed on the basis of HR-HPV status, macrophages were discriminated using CD68 and CD163 proteins as markers of pan-macrophage and M2 phenotype. The intensity of infiltrates was quantified by means of computer-assisted analysis in digital images of whole slides (virtual slides) separately in tumour tissue and stroma. Results: In HPV-positive patients, significantly more intense infiltration of both M1/M2 and M2 macrophages was found in the tumour stroma compared to HPV-negative patients. The infiltrates from both types of macrophages in the tumour tissue were less intense and did not differ between these groups. Intensive infiltration of CD68+ macrophages in the tumour front was associated with higher rate of nodal failures and a shorter nodal control in both HR-HPV groups. In the group of HR-HPV-negative patients, heavy infiltration of CD163+ macrophages was associated with significantly shorter: loco-regional control (LRC), metastasis-free survival and overall survival (OS). These parameters and prognosis in patients with scanty CD163+ infiltration were similar to favourable outcomes in HR-HPV-positive patients. The relative risk of local-regional recurrence, distant metastases and disease-related death in HR-HPV-negative patients with intense CD163+ infiltrates was, respectively, 4.7, 5.4 and 5.7 compared to patients with scanty infiltrates. Conclusions: Tumours with a positive HR-HPV status demonstrate intense infiltrations of total pool M1/M2 and M2 macrophages. In the group of HPV-negative patients, intensive M1/M2 macrophage infiltrates correlate with higher risk of nodal failures, and intensive M2 infiltrates are an adverse prognostic factor for LRC, metastasis-free survival and OS.

中文翻译：

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M2 样肿瘤相关巨噬细胞的浸润是人乳头瘤病毒阴性患者的不良预后因素，但不是人乳头瘤病毒阳性的口咽鳞状细胞癌

简介：具有高致癌潜力的人乳头瘤病毒 (HR-HPV) 是造成口咽部鳞状细胞癌一半以上的原因。尽管在诊断时疾病通常更晚期，但与 HR-HPV 阴性病例相比，该肿瘤的 HR-HPV 依赖性病例具有更好的预后。除了遗传和表观遗传因素外，这种更有利的疾病进程的原因还在于肿瘤微环境的参与，包括患者的免疫系统。巨噬细胞是构成肿瘤微环境的免疫活性细胞景观中最重要的元素之一。传统上，它们分为两组：具有 M1 表型的炎性巨噬细胞和称为 M2 表型巨噬细胞的肿瘤相关巨噬细胞。客观的：本研究的目的是分别研究 M1/M2 和 M2 表型的巨噬细胞浸润强度对口咽鳞状细胞癌 (OPSCC) 患者临床结果的影响，同时考虑到患者的 HR-HPV 状态。肿瘤。方法：该研究涉及 85 名 OPSCC 患者，其中使用双重检查算法确定肿瘤组织中的 HR-HPV 状态，包括通过 RT-PCR 方法检测病毒 DNA，随后通过免疫组织化学证实 P16INK4A 蛋白的生物学活性过度表达。在根据 HR-HPV 状态形成的每个组中，使用 CD68 和 CD163 蛋白作为泛巨噬细胞和 M2 表型的标志物来区分巨噬细胞。浸润的强度通过计算机辅助分析在肿瘤组织和基质中单独的整个载玻片（虚拟载玻片）的数字图像中量化。结果：在 HPV 阳性患者中，与 HPV 阴性患者相比，在肿瘤基质中发现 M1/M2 和 M2 巨噬细胞浸润明显更强烈。来自肿瘤组织中两种类型巨噬细胞的浸润不那么强烈，并且在这些组之间没有差异。在两个 HR-HPV 组中，肿瘤前沿 CD68+ 巨噬细胞的密集浸润与较高的淋巴结失败率和较短的淋巴结控制相关。在 HR-HPV 阴性患者组中，CD163+ 巨噬细胞的重度浸润与显着缩短相关：局部区域控制 (LRC)、无转移生存期和总生存期 (OS)。CD163+ 浸润少的患者的这些参数和预后与 HR-HPV 阳性患者的良好结果相似。与浸润很少的患者相比，具有强烈 CD163+ 浸润的 HR-HPV 阴性患者局部区域复发、远处转移和疾病相关死亡的相对风险分别为 4.7、5.4 和 5.7。结论：具有阳性 HR-HPV 状态的肿瘤表现出总池 M1/M2 和 M2 巨噬细胞的强烈浸润。在 HPV 阴性患者组中，密集的 M1/M2 巨噬细胞浸润与较高的淋巴结失败风险相关，而密集的 M2 浸润是 LRC、无转移生存和 OS 的不良预后因素。与浸润很少的患者相比，具有强烈 CD163+ 浸润的 HR-HPV 阴性患者局部区域复发、远处转移和疾病相关死亡的相对风险分别为 4.7、5.4 和 5.7。结论：具有阳性 HR-HPV 状态的肿瘤表现出总池 M1/M2 和 M2 巨噬细胞的强烈浸润。在 HPV 阴性患者组中，密集的 M1/M2 巨噬细胞浸润与较高的淋巴结失败风险相关，而密集的 M2 浸润是 LRC、无转移生存和 OS 的不良预后因素。与浸润很少的患者相比，具有强烈 CD163+ 浸润的 HR-HPV 阴性患者局部区域复发、远处转移和疾病相关死亡的相对风险分别为 4.7、5.4 和 5.7。结论：具有阳性 HR-HPV 状态的肿瘤表现出总池 M1/M2 和 M2 巨噬细胞的强烈浸润。在 HPV 阴性患者组中，密集的 M1/M2 巨噬细胞浸润与较高的淋巴结失败风险相关，而密集的 M2 浸润是 LRC、无转移生存和 OS 的不良预后因素。具有阳性 HR-HPV 状态的肿瘤表现出总池 M1/M2 和 M2 巨噬细胞的强烈浸润。在 HPV 阴性患者组中，密集的 M1/M2 巨噬细胞浸润与较高的淋巴结失败风险相关，而密集的 M2 浸润是 LRC、无转移生存和 OS 的不良预后因素。具有阳性 HR-HPV 状态的肿瘤表现出总池 M1/M2 和 M2 巨噬细胞的强烈浸润。在 HPV 阴性患者组中，密集的 M1/M2 巨噬细胞浸润与较高的淋巴结失败风险相关，而密集的 M2 浸润是 LRC、无转移生存和 OS 的不良预后因素。