HUMAN PAPILLOMAVIRUS EXPRESSION AND ANTIGENICITY

我们的总体目标是确定对hpv感染的免疫反应的特征。 用于两个不同的目的：第一，识别符合以下条件的血清学标记 与a)HPV感染相关；b)预防 肛门生殖器癌的发展或扩散；或c)可预测预后的 侵袭或转移的指标；第二，了解免疫 对人乳头瘤病毒的反应以设计免疫治疗策略。我们开发了一种 第一代试剂和技术允许最初的 人乳头瘤病毒6型和16型蛋白抗体的鉴定 人类血清。细菌表达的融合蛋白在Western中的应用 印迹分析在研究E7抗体方面特别有用。 蛋白。我们还使用了酵母菌表达的E7在ELISA中，并将 继续研究对该肿瘤相关抗原的反应。最近我们 使用痘苗病毒重组体产生了HPV衣壳蛋白，并表明 主要衣壳蛋白L1，无论有没有L2，都会自我组装形成 在结构上与正宗病毒粒子相同的衣壳。我们有 用抗L1单抗建立夹心ELISA法 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(S)。多数 有趣的是，这些抗体与该病的病史显著相关 人乳头瘤病毒感染。一个重要的目标将是确定衣壳是否 抗体与肛门癌有关。我们已经确立了 合作开始检查细胞介导的免疫反应 人乳头瘤病毒感染与宿主人类白细胞抗原基因在免疫中的作用 回应。这一资助期的目标是继续审查 血清标志物与肛门发育的关系 恶性肿瘤，并扩大我们的研究以寻找T细胞反应和 以确定我们人群中的人类白细胞抗原类型。

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.