ANALYSIS OF PAPILLOMAVIRUS

乳头瘤病毒（pv）感染动物和人的上皮，通常在感染部位诱导良性增殖。然而，人类生殖器病变的恶性进展与某些HPV类型，最常见的是HPV 16之间有很强的联系。我们已经为HPV 16和其他由L1主衣壳蛋白或L1加L2（次要衣壳蛋白）组成的pv生成了病毒样颗粒（VLPs）。在动物模型中，经肠外注射纯化的VLPs可诱导高滴度的中和抗体和对实验性攻击的保护。基于这些结果，我们已经验证了GMP级VLP，并且最近完成了HPV16 VLP疫苗的一期临床试验。疫苗接种者，即使是那些在没有佐剂的情况下接种疫苗的人，也能产生高滴度的HPV16伪病毒粒子中和抗体，并且只有轻微的副作用。第二阶段试验目前正在进行中。我们也在开发备选疫苗。为了增加基于vlp的疫苗的治疗潜力，我们将非结构性HPV蛋白作为L2融合蛋白加入到vlp中。接种HPV16 E7嵌合VLP产生CD8限制性T细胞反应，保护小鼠免受表达E7的肿瘤细胞系的肿瘤攻击，并诱导已建立的肿瘤消退。为了增加粘膜抗体反应，我们在小鼠中测试了经鼻滴注纯化的VLPs。与肠外接种VLPs相比，该方案在女性生殖道中诱导分泌IgA和转体IgG。中和抗体在鼻内接种的小鼠整个发情周期中检测到，但在肠外接种的小鼠中没有检测到。为了打破b细胞的耐受性并诱导抗体对自身蛋白的反应，我们用自身多肽取代了VLP的免疫显性表位。具体来说，我们设计了一种L1蛋白，在颗粒表面显示了小鼠CCR5的第一个外环。用这些颗粒接种的小鼠诱导出高滴度的抗体，这些抗体结合细胞表面的CCR5并阻断培养细胞的HIV感染。用含有猴/人CCR5肽的类似颗粒接种的猕猴也能产生抗CCR5 IgG。这些发现提示了一种诱导自身抗体的通用方法，具有许多基础和应用研究的应用价值。-癌症预防，免疫应答，乳头瘤病毒，疫苗设计，病毒-细胞相互作用，艾滋病，抗体，自身抗体，癌症疫苗，CCR5，宫颈癌，宫颈发育不良，HIV， HPV，人类逆转录病毒疫苗，-人体受试者和人体组织，液体，细胞等。

Papillomaviruses (PVs) infect the epithelia of animals and man where they generally induce benign proliferation at the site of infection. However, there is a strong association between malignant progression of human genital lesions and certain HPV types, most frequently HPV 16. We have generated virus-like particles (VLPs) for HPV 16 and other PVs that consist of the L1 major capsid protein or L1 plus L2, the minor capsid protein. Parenteral injection of purifed VLPs induced high titers of neutralizing antibodies and protection from experimental challenge in animal models. Based upon these results, we have validated GMP grade VLPs and have recently completed a phase 1 clinical trial of an HPV16 VLP vaccine. Vaccinees, even those vaccinated in the absence of adjuvant, produced high titers of HPV16 psuedovirion neutralizing antibodies and reported only minor side effects. A phase 2 trial is currently in progress.We are also developing alternative vaccine candidates. To increase the therapeutic potential of a VLP-based vaccine, we have incorporated non-structural HPV proteins into the VLPs as L2 fusion proteins. Vaccination with an HPV16 E7 chimeric VLP generated a CD8 restricted T cell response that protected mice from tumor challenge using an E7 expressing tumor line and also induced regression of established tumors. To increase mucosal antibody responses, we have tested intranasal instillation of purified VLPs in mice. In contrast to parenteral inoculation of VLPs, this protocol elicited both secretory IgA and transudated IgG in the female genital tract. Neutralizing antibodies were detected throughout the estus cycle in intranasally vaccinated mice, but not in those vaccinated parenterally.To break B-cell tolerance and induce antibody responses to a self-protein, we replaced an immunodominant epitope of a VLP with a self polypeptide. Specifically, we engineered a L1 protein that displayed the first external loop of mouse CCR5 on the surface of particles. Mice vaccinated with these particles induced high titers of antibodies that bound cell surface CCR5 and blocked HIV infection of culutred cells. Macaques vaccinated with similar particles displaying macaque/human CCR5 peptide also produced anti-CCR5 IgG. These findings suggest a general method for inducing auto-antibodies, with many basic and applied reasearch applications. - Cancer Prevention, immune response, Papillomaviruses, vaccine design, Virus-Cell Interactions, AIDS, Antibody, autoantibodies, Cancer vaccines, CCR5, cervical cancer, cervical dysplasia, HIV, HPV, vaccine against human retroviruses, - Human Subjects & Human Tissues, Fluids, Cells, etc.