PAPILLOMA VLP IMMUNIZATION AGAINST HPV-TRANSFORMED CELLS

针对 HPV 转化细胞的乳头状瘤 VLP 免疫

• 批准号: 6103360
• 负责人: WIJBE MARTIN KAST
• 金额: $ 15.12万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-14 至 2000-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6103360
• 关键词: B lymphocyte; antigen presenting cell; cell transformation; cytokine; cytotoxic T lymphocyte; fibroblasts; genetically modified animals; human papillomavirus; human tissue; immunization; keratinocyte; laboratory mouse; neoplasm /cancer immunology; neoplasm /cancer vaccine; neoplastic growth; neutralizing antibody; recombinant proteins; tissue /cell culture; tumor antigens; virus protein; virus related neoplasm /cancer; viruslike particle

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.

人乳头瘤病毒 (HPV) 与宫颈癌的病因有关 癌症。 95%以上的宫颈癌中均检测到 HPV DNA 其中 60% 是高危 HPV 16 型 (HPV16)。这 HPV16的E6和E7基因在人类中选择性保留和表达 癌并且足以使两种小鼠的转化 成纤维细胞和原代人角质形成细胞。这使得基本 小鼠免疫学实验可以推广到人类。高的 危险 HPV 感染肛门生殖器上皮的基底层 它们在其中复制的区域。由于这些复制条件不能 但经典疫苗的开发一直在体外进行模仿 由于无法获得足够数量的完整 HPV16 而受到阻碍 病毒体。然而，当 L1（病毒的主要病毒体蛋白）结束时 在真核细胞中表达，它组装成病毒样颗粒 （VLP）。表达病毒其他蛋白的重组 VLP，例如 E6和E7可以制作并将用于研究它们的预防和 治疗潜力是该提案的长期目标。 我们将实现以下具体目标： 1) 确定是否以及如何 VLP 免疫有效预防 HPV16 诱导的生长 小鼠体内的肿瘤。 2) 确定重组的治疗潜力 荷瘤小鼠体内的 VLP。 3) 确定重组的潜力 VLP 在人类 T 细胞培养物中诱导 HLA 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。为了实现这些 目标将使用以下方法：1) 对小鼠进行免疫 使用 VLP 并用 HPV16 诱导的肿瘤细胞进行攻击，特异性 VLP 中的中和抗体、T 细胞反应和肿瘤生长 将分析免疫小鼠。 2) 荷瘤小鼠的治疗 VLP 的局部或全身接种。 3) 人HPV特异性T细胞 人体外周血的体外免疫会诱导反应 抗原呈递细胞被 VLP 假性感染的血细胞。 4） 针对其他肿瘤相关抗原或细胞因子重组的 VLP 将被 分析其免疫效果并与其他疫苗接种进行比较 策略。这一综合分析将揭示 VLP 诱导预防性和/或治疗性 T 细胞介导的免疫 针对病毒诱发的肿瘤（如 HPV16 诱发的宫颈癌）的反应 在患者中。