HUMAN PAPILLOMAVIRUS EXPRESSION AND ANTIGENICITY

我们的总体目标是描述对HPV感染的免疫反应 为了两个不同的目的：第一，确定血清学标记， 与a）HPV感染; B）预防HPV感染的保护作用相关。 肛门生殖器癌的发展或扩散;或c）具有预后性 浸润或转移的指标;第二，了解免疫 设计免疫策略。我们开发了一个 第一代试剂和技术， 鉴定各种HPV 6和16蛋白的抗体， 人血清细菌表达的融合蛋白在Western blot中的应用 印迹分析在研究E7抗体方面特别有用 蛋白我们还在ELISA中使用了酵母表达的E7， 继续研究对肿瘤相关抗原的反应。最近我们 使用牛痘重组体生产HPV衣壳，并且已经表明， 主要衣壳蛋白，L1，有或没有L2，自组装形成 与真正的病毒粒子结构相同的衣壳。我们有 开发了一种夹心ELISA，使用L1的单克隆抗体捕获 衣壳颗粒，并在人类血清中检测到抗体， 识别天然HPV颗粒上的构象表位。最 有趣的是，这些抗体与 HPV感染。一个重要的目标将是确定是否衣壳 抗体与肛门生殖器癌有关。我们建立 合作，开始检查细胞介导的免疫反应， HPV感染与宿主HLA基因型在免疫应答中的作用 反应本供资期的目标是继续审查 血清学标志物与肛门生殖器发育的关系 恶性肿瘤，并扩大我们的研究，寻找T细胞反应， 来描述人类的HLA类型

Our overall goal is to characterize the immune response to HPV infection for two distinct purposes: first, to identify serological markers that are correlated with a) HPV infection; b) protection in preventing the development or spread of anogenital cancers; or c) that are prognostic indicators of invasion or metastasis; and second, to understand the immune response to HPV to design immunotherapeutic strategies. We developed a first generation of reagents and techniques that allowed the initial identification of antibodies to a variety of HPV 6 and 16 proteins in human sera. The use of bacterially expressed fusion proteins in Western blot assays was particularly informative in studying antibodies to the E7 protein. We have also used yeast-expressed E7 in an ELISA and will continue to study responses to that tumor associated antigen. Recently we produced HPV capsids using vaccinia recombinants and have shown that the major capsid protein, L1, with or without L2, self-assembles to form capsids that are structurally identical to authentic virions. We have developed a sandwich ELISA using monoclonal antibodies to L1 to capture the capsid particles, and have detected antibodies in human sera that recognize a conformational epitope(s) on the native HPV particle. Most interestingly these antibodies correlate significantly with a history of HPV infection. An important goal will be to determine whether capsid antibodies are related to anogenital cancers. We have established collaborations to begin to examine the cell mediated immune response to HPV infection and the role of the host HLA genotype in the immune response. The goals of this funding period are to continue to examine the relationship between serological markers and the development of anogenital malignancies, and to expand our studies to look for T cell responses and to characterize the HLA type in our populations.