Abstract：
Previous studies reported the association between Epstein-Barr virus (EBV) and cervical squamous cell carcinoma (CSCC), but its infection pattern and clinical significance unclear. This study aimed to comprehensively investigate the infection pattern, clinicopathology, outcomes, and immunology of this entity in central China. We evaluated a total of 104 untreated CSCC tumor tissue specimens using in situ hybridization for EBV-encoded small RNAs (EBERs), and by employing flowcytometry fluorescence hybridization for human papillomavirus (HPV) genotyping. The expression of EBV latency proteins and immune biomarkers was evaluated and quantified by immunohistochemistry. EBERs transcripts were detected in 21 (20.2%) cases overall (in malignant epithelial cells of 13 cases and in lymphocytes of 8 cases). EBV belonged to latency type I infection in CSCC. The high-risk (HR)-HPV was detected in all of EBV-positive CSCC, and the difference of detection rate of HR-HPV was significant when compared with EBV-negative CSCC (p = 0.001). The specific clinicopathology with increased frequency of advanced clinical stages, tumor-positive lymph nodes, neural invasion, and increased infiltration depth (all p value < 0.05) were observed in cases with EBV. However, EBV infection was found to have no impact on prognosis of patients with CSCC. Increased densities of forkhead box P3 (FoxP3)+-tumor infiltrating lymphocytes (TILs) (p = 0.005) and cytotoxic T-lymphocyte-associated protein 4 (CTLA-4)+-TILs (p = 0.017) and higher expression of programmed cell death-1 (PD-1) (p = 0.002) and programmed cell death-1 ligand 1 (PD-L1) (p = 0.040) were associated with EBV latent infection in CSCC, and these immunological changes were more likely to be associated with the infection in lymphocytes rather than tumor cells. Moreover, in patients with HPV-positive CSCC, similar significant differences were still found. In conclusions, EBV-positive CSCC may have specific infection pattern and clinicopathology and can exhibit an immunosuppressive microenvironment dominated by Treg cells aggregation and immune checkpoint activation.
摘要：
先前的研究报道了EB病毒（EBV）与宫颈鳞状细胞癌（CSCC）之间的关联，但其感染规律和临床意义尚不清楚。本研究旨在全面调查感染模式，临床病理,结果，和中国中部这个实体的免疫学。我们使用原位杂交评估了总共104个未经治疗的CSCC肿瘤组织标本的EBV编码的小RNA（EBERs），并采用流式细胞仪荧光杂交技术进行人乳头瘤病毒(HPV)基因分型。通过免疫组织化学评估和定量EBV潜伏期蛋白和免疫生物标志物的表达。在整体21例（20.2％）中检测到EBERs转录本（在13例恶性上皮细胞和8例淋巴细胞中）。EBV在CSCC中属于潜伏期I型感染。在所有EBV阳性的CSCC中均检测到高危型（HR）-HPV，与EBV阴性CSCC相比，HR-HPV检出率差异有统计学意义（p=0.001）。晚期临床分期频率增加的特定临床病理，肿瘤阳性淋巴结，神经入侵，在EBV病例中观察到浸润深度增加（所有p值<0.05）。然而,发现EBV感染对CSCC患者的预后没有影响。叉头盒P3（FoxP3）+-肿瘤浸润淋巴细胞（TIL）（p=0.005）和细胞毒性T淋巴细胞相关蛋白4（CTLA-4）+-TIL（p=0.017）的密度增加以及程序性细胞死亡-1（PD-1）（p=0.002）和程序性细胞死亡-1配体1（PD-L1）（p=0.040）的表达增加与CSCC潜伏EBV感染有关，这些免疫学变化更可能与淋巴细胞而不是肿瘤细胞的感染有关。此外，在HPV阳性CSCC患者中，仍然发现类似的显著差异。在结论中，EBV阳性CSCC可能具有特定的感染模式和临床病理，并且可以表现出由Treg细胞聚集和免疫检查点激活主导的免疫抑制微环境。
