Department of Molecular Virology, Faculty of Biology, University of A. Mickiewicz, Poznan, Poznan.
Viral Immunol. 2010 Aug;23(4):415-23. doi: 10.1089/vim.2010.0014.
Infection with human papillomaviruses (mostly HPV6 and HPV11) may lead to recurrent respiratory papillomatosis (RRP), a chronic disease affecting 2-4/100,000 people. Papillomas have to be removed surgically so patients can breathe normally. Papillomas often grow back and some patients are subjected to a number of operations. In general, asymptomatic HPV-positive people have low levels of antiviral antibodies in their sera, as the human humoral response is weak due to HPV's biology. In patients suffering from RRP who have undergone multiple surgeries, a blood-epithelium barrier breach stimulates the production of anti-HPV antibodies. Our study's aim was to produce HisTag-HPV11-L1 major capsid protein in E. coli cells, and to purify it. We also sought to detect anti-HPV11-L1 antibodies in antisera obtained from RRP patients using ELISA. Clinical samples were collected from 47 patients with RRP (antisera and papillomas), and from 32 controls (sera and oral swabs), from the Wielkopolska region of Poland. Antisera and control sera were used to coat microplates, HisTag-HPV11-L1 antigen was applied, and antibody-antigen complexes were detected by anti-HisTag monoclonal antibody in an ELISA assay. Simultaneously, total cellular DNA was extracted from papillomas and oral squamous cells obtained from controls. All DNA samples were screened for HPV DNA using MY-PCR. All patients were HPV-positive (30% for HPV6 and 70% for HPV11). Statistically significant correlations were found between the amount of anti-HPV11-L1 antibodies in the sera of RRP patients and the number of surgical procedures they underwent. Although HPV virus-like particles are most often used for anti-HPV antibody detection, the ELISA method presented herein is another viable option for use in RRP patients.
人乳头瘤病毒（HPV）感染可能导致复发性呼吸道乳头瘤病（RRP），这是一种影响每 10 万人中 2-4 人的慢性疾病。乳头状瘤必须通过手术切除，以便患者能够正常呼吸。乳头状瘤经常会复发，一些患者需要进行多次手术。一般来说，无症状的 HPV 阳性者血清中抗病毒抗体水平较低，因为 HPV 的生物学特性导致人体体液免疫反应较弱。在接受过多次手术的 RRP 患者中，血液-上皮屏障的破裂会刺激抗 HPV 抗体的产生。我们的研究目的是在大肠杆菌细胞中产生 HisTag-HPV11-L1 主要衣壳蛋白，并对其进行纯化。我们还试图使用 ELISA 检测从 RRP 患者获得的抗血清中抗 HPV11-L1 抗体。从波兰大波兰地区的 47 例 RRP 患者（抗血清和乳头状瘤）和 32 例对照者（血清和口腔拭子）中收集临床样本。将抗血清和对照血清包被在微孔板上，应用 HisTag-HPV11-L1 抗原，然后通过 ELISA 检测抗 HisTag 单克隆抗体检测抗体-抗原复合物。同时，从对照者的口腔鳞状细胞和乳头状瘤中提取总细胞 DNA。使用 MY-PCR 对所有 DNA 样本进行 HPV DNA 筛查。所有患者均为 HPV 阳性（HPV6 占 30%，HPV11 占 70%）。在 RRP 患者的血清中抗 HPV11-L1 抗体的量与他们接受的手术次数之间发现了统计学上的显著相关性。虽然 HPV 病毒样颗粒最常用于抗 HPV 抗体检测，但本文提出的 ELISA 方法是 RRP 患者的另一种可行选择。
