von Witzleben Adrian, Currall Eve, Wood Oliver, Chudley Lindsey, Akinyegun Oluyemisi, Thomas Jaya, Bendjama Kaïdre, Thomas Gareth J, Friedmann Peter S, King Emma V, Laban Simon, Ottensmeier Christian H
CRUK and NIHR Experimental Cancer Medicine Center & School of Cancer Sciences, Faculty of Medicine, University of Southampton, Southampton, United Kingdom.
Department of Otorhinolaryngology, Head & Neck Surgery, University of Ulm, Ulm, Germany.
Front Oncol. 2021 Jan 26;10:591063. doi: 10.3389/fonc.2020.591063. eCollection 2020.
Human papillomavirus 16 (HPV16) is the main cause of oropharyngeal squamous cell carcinoma (OPSCC). To date, the links between HPV16 gene expression and adaptive immune responses have not been investigated. We evaluated the correlation of HPV16 DNA, RNA transcripts and features of adaptive immune response by evaluating antibody isotypes against E2, E7 antigens and density of tumor-infiltrating lymphocytes (TIL).
FFPE-tissue from 27/77 p16-positive OPSCC patients was available. DNA and RNA were extracted and quantified using qPCR for all HPV16 genes. The TIL status was assessed. Immune responses against E2 and E7 were quantified by ELISA (IgG, IgA, and IgM; 77 serum samples pre-treatment, 36 matched post-treatment).
Amounts of HPV16 genes were highly correlated at DNA and RNA levels. RNA co-expression of all genes was detected in 37% (7/19). E7 qPCR results were correlated with higher anti-E7 antibody (IgG, IgA) level in the blood. Patients with high anti-E2 IgG antibody (>median) had better overall survival (p=0.0311); anti-E2 and anti-E7 IgA levels had no detectable effect. During the first 6 months after treatment, IgA but not IgG increased significantly, and >6 months both antibody classes declined over time. Patients with immune cell-rich tumors had higher levels of circulating antibodies against HPV antigens.
We describe an HPV16 qPCR assay to quantify genomic and transcriptomic expression and correlate this with serum antibody levels against HPV16 oncoproteins. Understanding DNA/RNA expression, relationship to the antibody response in patients regarding treatment and outcome offers an attractive tool to improve patient care.
人乳头瘤病毒16型（HPV16）是口咽鳞状细胞癌（OPSCC）的主要病因。迄今为止，HPV16基因表达与适应性免疫反应之间的联系尚未得到研究。我们通过评估针对E2、E7抗原的抗体亚型以及肿瘤浸润淋巴细胞（TIL）的密度，来评估HPV16 DNA、RNA转录本与适应性免疫反应特征之间的相关性。
可获得27例/77例p16阳性OPSCC患者的福尔马林固定石蜡包埋组织（FFPE组织）。使用qPCR对所有HPV16基因进行DNA和RNA提取及定量分析。评估TIL状态。通过酶联免疫吸附测定（ELISA）（IgG、IgA和IgM；77份血清样本治疗前，36份匹配的治疗后样本）对针对E2和E7的免疫反应进行定量分析。
HPV16基因的数量在DNA和RNA水平上高度相关。在37%（7/19）的样本中检测到所有基因的RNA共表达。E7 qPCR结果与血液中较高的抗E7抗体（IgG、IgA）水平相关。抗E2 IgG抗体水平高（>中位数）的患者总生存期更好（p=0.0311）；抗E2和抗E7 IgA水平未显示出可检测到的影响。在治疗后的前6个月，IgA而非IgG显著增加，6个月后两种抗体类别均随时间下降。免疫细胞丰富的肿瘤患者针对HPV抗原的循环抗体水平更高。
我们描述了一种HPV16 qPCR检测方法，用于定量基因组和转录组表达，并将其与针对HPV16癌蛋白的血清抗体水平相关联。了解DNA/RNA表达、患者治疗及预后方面与抗体反应的关系，为改善患者护理提供了一个有吸引力的工具。
