PAPILLOMAVIRUS MODEL SYSTEMS FOR MICROBICIDES

人乳头瘤病毒（hpv）代表一些最常见的性传播疾病。已经确定了80多种HPV类型，其中几种类型与宫颈癌、阴道癌、阴茎癌、口腔癌和皮肤癌的发病密切相关。HPV DNA几乎存在于所有宫颈癌病例（93%）及其前驱病变中。目前具有乳头瘤病毒活性的杀微生物化合物尚未被描述。此外，还没有发表过关于生殖器乳头瘤病毒感染的动物模型的报告，用于测试阴道内配方和同时发生的性病感染。最近，一些体外和体内乳头瘤病毒感染模型已被证实，这使得筛选和测试潜在的抗乳头瘤病毒活性的杀微生物化合物和配方成为可能。本更新建议的目的是建立两个体外和两个体内乳头瘤病毒感染模型的有效性，用于检测杀微生物乳头瘤病毒化合物。我们建议使用的体外模型是(I) bpv -1诱导小鼠C127细胞的病灶形成，以及（ii）使用基于elisa的RT-PCR检测HPV E1 E4剪接mRNA转录物的HPV-11（和HPV-40）在体外短暂感染人A431上皮细胞。我们将使用体外模型筛选有效的抗乳头瘤病毒化合物，然后在两种动物模型系统中进行测试。其中包括使用感染性HPV-11和HPV-40病毒粒子的新型人类阴道异种移植动物模型系统，以及我们实验室最近开发的兔生殖器乳头瘤病毒模型。在本项目的前3年，我们已经确定了几种潜在的乳头瘤病毒杀微生物化合物，并开始表征生殖器乳头瘤病毒感染的体内模型。这些杀微生物剂（中和单克隆抗体和烷基硫酸盐）是第一批具有抗乳头瘤病毒活性的化合物。我们将继续对其进行表征，并使用阴道内（进入人类阴道异种移植物和完整的兔子阴道）的配方扩大测试，以获得体内疗效。

Human papillomaviruses (HPVs) represent some of the most common of the sexually transmitted disease. More than 80 HPV types have been identified and several types have been strongly implicated as causative agents in the onset of cervical vaginal, penile, oral and skin cancers. HPV DNA is present in virtually all cases (93 percent) of cervical cancer and its precursor lesions. Current microbicidal compounds with papillomavirus activity have not been described. In addition, there are not published reports of animal models of genital papillomavirus infections for testing intra-vaginal formulations and co-incidental STD infections. Recently, several in vitro and in vivo papillomavirus infectivity models have been documented that make feasible the screening and testing of microbicidal compounds and formulations for potential anti- papillomavirus activity. The purpose of this renewal proposal is to establish the validity of two in vitro and two in vivo papillomavirus infectivity models for the testing of microbicidal papillomavirus compounds. The in vitro models we propose to use are (I) BPV-1-induced focus formation of mouse C127 cells, and (ii) transient in vitro HPV-11 (and HPV-40) infection of human A431 epithelial cells using an ELISA-based RT-PCR detection of HPV E1 E4 spliced mRNA transcripts. We will use the in vitro models to screen for effective anti-papillomavirus compounds to be later tested in two animal model systems. These include a novel human vaginal xenograft animal model system using infectious HPV-11 and HPV-40 virions, and a rabbit genital papillomavirus model recently developed in our laboratory. During the first 3 years of this Program Project, we have identified several potential papillomavirus microbicidal compounds and have begun to characterize in vivo model of genital papillomavirus infections. These microbicidal agents (neutralizing monoclonal antibodies and alkyl sulfates) represent the first compounds with anti-papillomavirus activity. We will continue their characterizing and expand the testing using formulations delivered intra-vaginally (into human vaginal xenografts and into intact rabbit vaginas) for in vivo efficacy.