PAPILLOMA VLP IMMUNIZATION AGAINST HPV-TRANSFORMED CELLS

人类乳头瘤病毒(HPV)与宫颈病变有关 癌症。HPV DNA在95%以上的宫颈癌和 其中60%是由高危HPV16型(HPV16)引起的。这个 人乳头瘤病毒16型E6和E7基因在人体内选择性保留和表达 并足以转化为小鼠和小鼠 成纤维细胞和原代人角质形成细胞。这允许基本的 小鼠的免疫学实验可以外推到人类身上。高 高危HPV感染肛门生殖器上皮基底层 它们在其中复制的区域。由于这些复制条件不能 然而，在体外被模仿，经典疫苗的开发一直是 由于缺乏获得足够数量的完整HPV16而受阻 病毒粒子。然而，当L1，病毒的主要病毒粒子蛋白结束 在真核细胞中表达，它会组装成病毒样颗粒 (VLP)。表达病毒其他蛋白的重组VLP，如 E6和E7可以制成，并将用于研究它们的预防性和 治疗潜力，这是这项提议的长期目标。 我们将解决以下具体目标：1)确定是否以及如何 VLP免疫可有效阻止HPV16感染后的生长 小鼠体内的肿瘤。2)确定重组人的治疗潜力。 荷瘤小鼠的血管内皮细胞。3)确定重组人的潜能 VLP在人T细胞培养中诱导人类白细胞抗原限制性T细胞反应 体外和4)确定重组VLP的免疫潜力 表达其他肿瘤相关抗原或细胞因子。要实现这些目标 目标：将采用以下方法：1)免疫小鼠 用VLP和HPV16诱导的肿瘤细胞攻击，特异性 中和抗体、T细胞反应与VLP肿瘤生长 免疫的小鼠将被分析。2)荷瘤小鼠将接受以下治疗 局部或系统接种VLP。3)人HPV特异性T细胞 人体外周血细胞体外免疫可诱导免疫应答 带有抗原提呈细胞的血细胞假性感染VLP。4) 为其他肿瘤相关抗原或细胞因子重组的VLP将被 分析它们的免疫效果，并与其他疫苗进行比较 战略。这一综合分析将揭示出 VLP诱导预防性和/或治疗性T细胞介导的免疫 对病毒诱导的肿瘤如HPV16诱导的宫颈癌的反应 在病人身上。

Human papillomaviruses (HPVs) are implicated in the etiology of cervical cancer. HPV DNA is detected in more than 95% of cervical carcinomas and 60% of these are accounted for by the high risk HPV type 16 (HPV16). The E6 and E7 genes of HPV16 are selectively retained and expressed in human carcinomas and are sufficient for the transformation of both mouse fibroblasts and primary human keratinocytes. This allows fundamental immunological experiments in mice that can be extrapolated to humans. High risk HPVs infect the basal layers of the epithelium of the anogenital tract in which they replicate. Since these replication conditions can not yet be mimicked in vitro, the development of classical vaccines has been hampered by the lack of obtaining sufficient amounts of intact HPV16 virions. However, when L1, the major virion protein of the virus is over expressed in eukaryotic cells, it assembles into virus-like particles (VLPs). Recombinant VLPs that express other proteins of the virus such as E6 and E7 can be made and will be used to study their preventative and therapeutic potential which is the long-term objective of this proposal. We will address the following specific aims: 1) To determine if and how VLP immunization effectively prevents the outgrowth of HPV16 induced tumors in mice. 2) To determine the therapeutic potential of recombinant VLPs in tumor-bearing mice. 3) To determine the potential of recombinant VLPs to induce HLA-restricted T cell responses in human T cell cultures in vitro and 4) To determine the immunologic potential of recombinant VLPs expressing other tumor-associated antigens or cytokines. To achieve these aims the following methodology will be used: 1) mice will be immunized with VLPs and challenged with HPV16-induced tumor cells, specific neutralizing antibodies, T cell responses and tumor out growth in VLP immunized mice will be analyzed. 2) Tumor bearing mice will be treated by local or systemic inoculation of VLPs. 3) Human HPV specific T cell responses will be induced by in vitro immunization of human peripheral blood cells with antigen presenting cells pseudo-infected with VLPs. 4) VLPs recombinant for other tumor-associated antigens or cytokines will be analyzed for their immunizing effects and compared to other vaccination strategies. This combined analysis will shed light on the potential of VLPs to induce preventative and/or therapeutic T-cell mediated immune responses against virus induced tumors like HPV16 induced cervical cancer in patients.