HPV CAPSID ANTIBODIES

HPV感染的流行率估计是基于 在生殖器部位检测HPV DNA，这种方法不允许 区分原发性感染和复发性感染。能够使用 HPV抗体阳性作为感染的标志物受到了阻碍， 缺乏合适的HPV抗原来源。我们最初的努力， 蛋白质印迹中细菌表达的蛋白质提供了有力的证据， HPV血清抗体，但表明抗体反应， 构象表位可能是免疫应答的重要组成部分。 反应我室牛痘病毒研究进展 表达的衣壳在结构上与真正的HPV相同 病毒粒子使得ELISA的发展，检测“类型特异性” 与主要衣壳上的构象表位反应的抗体 蛋白质，L1.报告HPV感染的女性中HPV 1衣壳的血清阳性率 有足疣史者占89%。HPV 6血清阳性与 在几个人群中有当前或历史生殖器疣，HPV 6/11在生殖道和性伴侣数量中检测到DNA 在大学女性群体中。这些结果构成了 建议分析血清中的HPV 6型和16型抗体， 正在进行的队列：正在开始性活动的大学女性和 一群孕妇和她们的孩子。述纵向收集 将使我们能够研究外观的动力学， 抗衣壳抗体的消失，以提供 血清转化，检查IgM和IgG抗体的出现， 定义与开发相关的参数， 抗体应答，如病毒载量、病毒类型、年龄、性别、后遗症 感染和其他因素。L1基因将被克隆、测序， 从血清阴性但DNA阳性的受试者表达， 确定抗原变异是否可以解释缺乏 血清反应性将获取粘液分泌物以寻找证据 本地抗体。系统发育分析提供了深入了解 HPV类型的相关性。表达以下蛋白的痘苗病毒重组体 所有生殖器类型的主要衣壳蛋白， 或其他 遗传学上相关的类型，例如2和27，仅在 将构建在皮肤部位检测的衣壳， 用于确定是否可以开发HPV血清型系统。 交叉反应性将使用实验衍生血清进行评估， 阻断人类血清的实验随着新衣壳的产生， 将确定这些类型的血清阳性率。这些数据将是 了解HPV感染自然史的重要辅助手段 并将为开发和测试 HPV疫苗。

Estimates of the prevalence of HPV infection have been based on the detection of HPV DNA at genital sites, and this approach has not allowed a distinction between primary or recurrent infection. The ability to use HPV antibody positivity as a marker of infection has been hampered by the lack of a suitable source of HPV antigens. Our initial efforts using bacterially expressed proteins in Western blots provided firm evidence of HPV serum antibodies but suggested that antibodies reactive with conformational epitopes may be an important component of the immune response. The recent development in our laboratory of vaccinia virus expressed capsids that are structurally identical to authentic HPV virions has allowed the development of ELISAs that detect "type specific" antibodies reactive with a conformational epitope(s) on the major capsid protein, L1. Seropositivity to HPV 1 capsids among women reporting a history of foot warts was 89%. Seropositivity to HPV 6 was associated with current or historical genital warts in several populations, with HPV 6/11 DNA detected in the genital tract and with number of sexual partners in a population of college women. These results form the basis for proposing to analyze sera for antibodies to HPV types 6 and 16 in two ongoing cohorts: college women who are initiating sexual activity and a cohort of pregnant women and their babies. The longitudinal collection of sera will allow us to study the kinetics of appearance and disappearance of anti-capsid antibodies, to provide evidence of seroconversion, to examine the appearance of IgM and IgG antibodies and to define the parameters that are associated with the development of an antibody response such as viral load, viral type, age, gender, sequelae of infection and other factors. The L1 gene will be cloned, sequenced and expressed from subjects who are seronegative but DNA positive, to determine whether antigenic variants can explain the lack of seroreactivity. Mucosal secretions will be obtained to look for evidence of local antibodies. Phylogenetic analyses have provided insights into the relatedness of HPV types. Vaccinia virus recombinants expressing the major capsid proteins for all of the genital types and or other phylogenetically related types, e.g. 2 and 27 that have only been detected at cutaneous sites, will be constructed and the capsids will be used to determine whether a system of HPV serotypes can be developed. Cross-reactivity will be assessed using experimental derived sera and in blocking experiments with human sera. As new capsids are developed, seroprevalence to these types will be determined. These data will be an important adjunct for understanding the natural history of HPV infection and will provide critical information for the development and testing of HPV vaccines.