PAPILLOMA VLP IMMUNIZATION AGAINST HPV-TRANSFORMED CELLS

人乳头瘤病毒（HPV）与宫颈癌的病因有关， 癌HPV DNA在超过95%的宫颈癌中检测到， 其中60%是高危型HPV 16（HPV 16）。的 人乳头瘤病毒16型E6和E7基因在人中选择性保留和表达 癌，并足以转化两个小鼠 成纤维细胞和原代人角质形成细胞。这使得基础 小鼠的免疫学实验可以外推到人类。高 高危HPV感染肛门生殖器上皮的基底层， 在其中复制。由于这些复制条件不能 然而，在体外模拟，经典疫苗的发展已经 由于缺乏足够数量的完整HPV 16， 病毒粒子然而，当病毒的主要病毒体蛋白L1结束时， 在真核细胞中表达，它组装成病毒样颗粒 （VLP）。表达病毒的其他蛋白质的重组VLP， E6和E7可以制备并将用于研究其预防和治疗作用。 治疗潜力，这是本提案的长期目标。 我们将讨论以下具体目标：1）确定是否以及如何 VLP免疫有效地阻止了HPV 16诱导的生长， 小鼠的肿瘤2)为了确定重组的治疗潜力 荷瘤小鼠中的VLP。3)为了确定重组的潜力， VLP在人T细胞培养物中诱导HLA限制性T细胞应答 体外和4）确定重组VLP的免疫潜力 表达其它肿瘤相关抗原或细胞因子。实现这些 目的：1）免疫小鼠， 用VLP和用HPV 16诱导的肿瘤细胞攻击，特异性 VLP中的中和抗体、T细胞应答和肿瘤生长 将分析免疫的小鼠。2)荷瘤小鼠将通过 VLP的局部或全身接种。3)人HPV特异性T细胞 将通过体外免疫人外周血淋巴细胞诱导应答。 用VLP假感染抗原呈递细胞的血细胞。四、 针对其他肿瘤相关抗原或细胞因子的VLP重组将被用于治疗肿瘤。 分析其免疫效果，并与其他疫苗进行比较 战略布局这一综合分析将揭示 诱导预防性和/或治疗性T细胞介导的免疫应答的VLP 对病毒诱导的肿瘤（如HPV 16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.