Capsid-incorporation of HIV antigens as a novel adenovirus HIV vaccine approach

首席调查员/项目主任(最后、第一、中间)：库里尔、大卫、T. 研究及相关其他项目信息 1.*是否涉及人类受试者？M是的，L不是 1.a.如果是，对人类受试者 IRB的审查正在进行中吗？M是m否 IRB批准日期： 豁免编号：1 2 3 4 5 6 人类主题保证编号 2.*是否使用脊椎动物？L：是，我不是 2.a.如果是，脊椎动物 IACUC的审查正在进行中吗？L：是，我不是 IACUC批准日期： 动物福利保障编号A3255-01 3.*是否是专有/特权信息m是L否 是否包含在应用程序中？ 4.a.*这个项目对我有没有实际或潜在的影响？是L否 环境？ 4.B.如果是，请解释： 4.C.如果该项目对环境有实际或潜在的影响，是否已批准豁免或环境评估(EA)或 是否执行了环境影响报告书(EIS)？M是m否 4.D.如果是，请解释： 5.a.*这个项目是否涉及美国以外的活动或m是L否 与国际合作者建立伙伴关系？ 5.B.如果是，请列出以下国家/地区： 5.c.可选解释： 6.*项目摘要/摘要1235-项目__摘要__Curiel.pdf模拟类型：应用程序/pdf 7.*Project Narrative 4856-Project__Narrative_Curiel.pdf模仿类型：应用程序/pdf 8.参考书目和参考文献3112-Reference_List_Curiel.pdf笔记类型：应用程序/pdf 9.设施和其他资源2477-FACILITIES.pdf模拟类型：应用程序/pdf 10.设备1015-EQUIPMENT.pdf模拟类型：应用/pdf 追踪编号：其他信息第5页OMB编号：4040-0001 到期日：04/30/2008 首席调查员/项目主任(最后、第一、中间)：库里尔、大卫、T. 尽管Ad载体在疫苗应用方面具有许多潜在的优势，但充分利用现有的Ad疫苗 可能受到宿主抗病媒免疫反应的限制。具体地说，反广告体液免疫废除 后续给药的有效性的广告载体，混淆表达的编码 转基因，从而实际上限制了可能通过助推器效应而产生的收益。为了利用 针对Ad载体固有的抗原性，我们开发了一种基于掺入 免疫抗原表位直接进入Ad衣壳。这种新的范式是基于广告呈现的 作为衣壳的组成部分的抗原，而不是编码的转基因。免疫原性掺入 多肽进入Ad衣壳提供了潜在的优势。最值得注意的是，衣壳的处理 通过外源途径掺入的抗原应该会产生类似于 由天然的Ad衣壳蛋白引起的反应。此外，由于增强了抗广告衣壳应答 通过重复载体注射，针对作为Ad衣壳一部分的抗原表位的免疫应答 也应该通过重复给药来增加，从而允许增加。这些考虑因素 提示这种新的衣壳包被抗原的方法可能为实现基于Ad的 规避与Ad载体相关的主要限制的疫苗策略。 实现这一方法的关键是确定腺病毒中抗原的最佳构型 衣冠楚楚的背景。为此，我们已经建立了几项关键技术，使我们能够达到我们的 进球。特别是，我们已经开发出将异源多肽表位整合到 主要的Ad衣壳蛋白Hexon的表面暴露结构域。我们已经开始确定大小和 结构因素，预测这些域在六角体中的功能效用。此外，我们还开发了 应用低温电子显微镜(CryoEM)单粒子重建方法的方法使我们能够 探索衣壳结合的多肽定位，具有前所未有的亚纳米分辨率。基于 这些技术，我们将能够建立抗原地点/可获得性之间的关键关联 在衣壳背景和疫苗效力方面。 在这些已建立的可行性的基础上，我们假设广告载体可以用新的 衣壳结合的抗原，可以在动物模型中用作艾滋病毒的疫苗制剂。低温电磁制导 将应用衣壳设计来开发具有最佳抗HIV免疫的优化载体。我们设想 我们提议的结构研究将为体外分析和生物学研究提供补充信息 因此，这将使我们能够理解所结合的艾滋病毒表位的功能决定因素。 项目说明第6页

Principal Investigator/Program Director (Last, first, middle): Curiel, David, T. RESEARCH & RELATED Other Project Information 1. * Are Human Subjects Involved? m Yes l No 1.a. If YES to Human Subjects Is the IRB review Pending? m Yes m No IRB Approval Date: Exemption Number: 1 2 3 4 5 6 Human Subject Assurance Number 2. * Are Vertebrate Animals Used? l Yes m No 2.a. If YES to Vertebrate Animals Is the IACUC review Pending? l Yes m No IACUC Approval Date: Animal Welfare Assurance Number A3255-01 3. * Is proprietary/privileged information m Yes l No included in the application? 4.a.* Does this project have an actual or potential impact on m Yes l No the environment? 4.b. If yes, please explain: 4.c. If this project has an actual or potential impact on the environment, has an exemption been authorized or an environmental assessment (EA) or environmental impact statement (EIS) been performed? m Yes m No 4.d. If yes, please explain: 5.a.* Does this project involve activities outside the U.S. or m Yes l No partnership with International Collaborators? 5.b. If yes, identify countries: 5.c. Optional Explanation: 6. * Project Summary/Abstract 1235-Project__Summary__Curiel.pdf Mime Type: application/pdf 7. * Project Narrative 4856-Project__Narrative_Curiel.pdf Mime Type: application/pdf 8. Bibliography & References Cited 3112-Reference_List_Curiel.pdf Mime Type: application/pdf 9. Facilities & Other Resources 2477-FACILITIES.pdf Mime Type: application/pdf 10. Equipment 1015-EQUIPMENT.pdf Mime Type: application/pdf Tracking Number: Other Information Page 5 OMB Number: 4040-0001 Expiration Date: 04/30/2008 Principal Investigator/Program Director (Last, first, middle): Curiel, David, T. Despite the many potential advantages of Ad vectors for vaccine application, full utility of current Ad vaccines may be limited by the host anti-vector immune response. Specifically, the anti-Ad humoral immunity abrogates the effectiveness of subsequent administrations of the Ad vector, confounding expression of the encoded transgene, and thus practically restricting the gains that might be accrued via booster effect. In order to exploit the inherent antigenicity of the Ad vector we have developed a vaccination approach based on incorporation of the immunizing antigen epitope directly into the Ad capsid. This novel paradigm is based upon Ad presenting the antigen as a component of the capsid rather than an encoded transgene. Incorporation of immunogenic peptides into the Ad capsid offers potential advantages. Most noteworthy, the processing of the capsid incorporated antigen via the exogenous pathway should result in a strong humoral response akin to the response provoked by native Ad capsid proteins. In addition, since anti-Ad capsid responses are augmented by repeated vector administration, immune responses against antigenic epitopes that are part of the Ad capsid should be augmented by repeated administration as well, thus allowing boosting. These considerations suggest that this novel capsid-incorporated antigen approach may offer exciting potentials to realize Ad-based vaccine strategies that circumvent the major limitations associated with Ad vectors. Critical to the realization of this approach is to define the optimal configuration of antigen in the adenoviral capsid context. To this end, we have established several key technologies that will enable us to reach our goal. In particular, we have developed the means to incorporate heterologous peptide epitopes within the surface-exposed domains of the major Ad capsid protein hexon. We have begun to determine the size and structural factors that predicate functional utility of these domains in the hexon. In addition, we have developed the means to apply cryoelectron microscopy (cryoEM) single particle reconstruction methods to allow us to explore the capsid-incorporated peptide localization with unprecedented, subnanometer resolution. Based on these technologies, we will be able to establish the critical correlates between antigen locale/accessibility within the capsid context and vaccine efficacy. On the basis of these established feasibilities, we hypothesize that Ad vectors can be created with novel capsid-incorporated antigens that can serve as vaccine agents against HIV in animal models. CryoEM-guided capsid design will be applied to develop an optimized vector with optimal anti-HIV immunization. We envision that our proposed structural studies will provide complementary information to in vitro assays and biological readouts and thereby will enable us to understand the functional determinants of incorporated HIV epitopes. Project Description Page 6