MINIGENE VACCINATION WITH EARLY PRESENTED VIRAL PROTEINSAIDS RELATED RESEARCH

早期呈现病毒蛋白的 MINIGENE 疫苗接种艾滋病相关研究

• 批准号: 8358214
• 负责人: David I Watkins
• 金额: $ 38.13万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8358214
• 关键词: Adenovirus Vector; Adenoviruses; Animals; CD4 Positive T Lymphocytes; CD8B1 gene; DNA; Electroporation; Epitopes; Equilibrium; Flow Cytometry; Frequencies; Funding; Gagging; Genetic Services; Goals; Grant; HIV; Human; Immune; Immune response; Immunodominant Epitopes; Immunology; Infection; Interleukin-12; Macaca; Macaca mulatta; Mutate; National Center for Research Resources; Plasmids; Primates; Principal Investigator; Proteins; PubMed; Publications; Recombinants; Regimen; Research; Research Infrastructure; Resources; SIV; Series; Services; Source; System; T cell response; T-Lymphocyte; Techniques; Testing; Time; United States National Institutes of Health; Vaccinated; Vaccination; Vaccines; Viral; Viral Load result; Viremia; Wisconsin; cost; design; immunogenicity; instrument; novel; novel strategies; novel vaccines; plasmid DNA; rBCG; response; vector; vector vaccine; virology

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Objective: To study a new vaccine approach to HIV. Immune epitope dominance results in one or a few epitopes comprising the greatest part of the immune response to SIV infection and vaccination. We know that some of these dominant epitopes are not useful in controlling SIV because they either mutate rapidly (Mamu-A*01 Tat SL8) or simply do not control viremia on their own (Mamu-A*01 Gag CM9). It may be that subdominant immune responses are more efficacious than dominant immune responses, but are at low frequency. Therefore, we have designed a series of Minigenes, which are regions of SIV proteins that encode no more than one immunodominant epitope. By separating immunodominant epitopes from subdominant epitopes, we hope to allow increased frequency of the subdominant epitopes resulting in greater breadth of the immune response. In addition, while we have had successful vaccine trials in rhesus macaques with DNA/Ad5 as vaccine vectors, adenovirus is now less favored as a vaccine vector in humans due to widespread prior infection with adenovirus and due to the poor results obtained in the recent Merck STEP trial using this vector. Therefore, at the same time that we are exploring the minigene concept, we are also exploring the use of alternate vectors. We will put minigenes into rBCG, rShigella, rYF-17D and possibly other vectors, vaccinate macaques and challenge them to assess the immunogenicity of these new approaches. PROGRESS: Following up on our previous publication (see Novel, Logical grant) we have started a new vaccine/challenge study with the goal of enhancing breadth of Gag, Vif and Nef, as well as to test new vectors and delivery systems. We are just finished with the vaccinations and will begin challenging in February 2011. In this study, we inserted six minigenes of Gag, two minigenes of Vif and the core region of Nef into DNA plasmids, recombinant YF-17D vectors and Ad5 vectors. These minigenes were optimized for expression by our colleagues at IAVI. We utilized several different vaccine strategies to compare use if IL-12 during vaccination, use of rYF as either a boost or a prime and comparison of DNA prime/Ad5 boost to rYF prime/Ad5 boost. For the DNA primes, we used a new technique called DNA electroporation, which greatly enhanced immune responses to the sequences encoded in the DNA plasmids. In one group, these responses were further enhanced by the addition of a plasmid encoding IL-12. We found that these immune responses were greatly biased toward CD4+ T cell responses. Subsequent rYF and/or rAd5 boosts resulted in a more balanced immune response, with the result that we have robust immune responses in both CD4+ and CD8+ T cells compartments. We also primed with rYF, then boosted with rAd5, because colleagues have shown that rYF acts more like a prime than a boost. Comparison of the responses engendered by this regimen confirmed that while the responses observed after rYF prime are low, the responses seen after boosting with rAd5 are much greater than that observed with rAd5 alone. Now that these vaccinations are completed, we will start challenging animals in February 2011. This research used WNPRC Animal Services, Genetics Services, and Immunology & Virology Services. This grant also utilized the resources of the MHC typing facility and Immunology and Virology Services (viral load determination, tetramers, flow cytometry instrument utilization, Elispots). PUBLICATION: Sacha JB, Buechler MB, Newman LP, Reed J, Wallace LT, Loffredo JT, Wilson NA, Watkins DI. Simian immunodeficiency virus-specific CD8+ T cells recognize Vpr- and Rev-derived epitopes early after infection. J Virol. 2010 Oct;84(20):10907-12. Epub 2010 Aug 4. PubMed PMID: 20686015; PubMed Central. PMCID: PMC2950557.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 目的：研究艾滋病毒的新疫苗方法。 免疫表位优势导致一个或几个表位构成对 SIV 感染和疫苗接种的免疫反应的大部分。我们知道，其中一些显性表位在控制 SIV 方面无用，因为它们要么快速突变 (Mamu-A*01 Tat SL8)，要么根本无法自行控制病毒血症 (Mamu-A*01 Gag CM9)。次显性免疫反应可能比显性免疫反应更有效，但频率较低。因此，我们设计了一系列小基因，它们是编码不超过一个免疫显性表位的 SIV 蛋白区域。通过将免疫显性表位与次显性表位分开，我们希望能够增加次显性表位的频率，从而产生更大范围的免疫反应。 此外，虽然我们已经在恒河猴中以 DNA/Ad5 作为疫苗载体进行了成功的疫苗试验，但由于腺病毒先前广泛感染以及最近默克公司获得的不佳结果，腺病毒现在不太受欢迎作为人类疫苗载体。使用该载体进行 STEP 试验。因此，我们在探索小基因概念的同时，也在探索替代载体的使用。我们将把小基因放入 rBCG、rShigella、rYF-17D 和可能的其他载体中，对猕猴进行疫苗接种并挑战它们以评估这些新方法的免疫原性。 进步： 继我们之前的出版物（参见小说、逻辑资助）之后，我们开始了一项新的疫苗/挑战研究，目标是增强 Gag、Vif 和 Nef 的广度，并测试新的载体和递送系统。我们刚刚完成疫苗接种，将于 2011 年 2 月开始挑战。 在本研究中，我们将Gag的6个小基因、Vif的2个小基因和Nef的核心区域插入DNA质粒、重组YF-17D载体和Ad5载体中。我们 IAVI 的同事对这些小基因的表达进行了优化。我们利用几种不同的疫苗策略来比较疫苗接种期间 IL-12 的使用、rYF 作为加强剂或初免剂的使用以及 DNA 初免/Ad5 加强剂与 rYF 初免/Ad5 加强剂的比较。对于 DNA 引物，我们使用了一种称为 DNA 电穿孔的新技术，该技术大大增强了对 DNA 质粒中编码的序列的免疫反应。在一组中，通过添加编码 IL-12 的质粒进一步增强了这些反应。我们发现这些免疫反应极大地偏向于 CD4+ T 细胞反应。随后的 rYF 和/或 rAd5 加强导致更平衡的免疫反应，结果是我们在 CD4+ 和 CD8+ T 细胞区室中都有强大的免疫反应。我们还用 rYF 引发，然后用 rAd5 增强，因为同事已经表明，rYF 的作用更像是引发而不是增强。对该方案产生的反应的比较证实，虽然rYF初免后观察到的反应较低，但用rAd5加强后观察到的反应比单独用rAd5观察到的反应大得多。现在这些疫苗接种已经完成，我们将于 2011 年 2 月开始对动物进行挑战。 这项研究使用了 WNPRC 动物服务、遗传学服务以及免疫学和病毒学服务。这笔赠款还利用了 MHC 分型设施以及免疫学和病毒学服务的资源（病毒载量测定、四聚体、流式细胞术仪器利用、Elispots）。 发布： Sacha JB、Buechler MB、Newman LP、Reed J、Wallace LT、Loffredo JT、Wilson NA、 沃特金斯 DI。猿猴免疫缺陷病毒特异性 CD8+ T 细胞识别 Vpr- 和感染后早期Rev衍生的表位。 J·维罗尔. 2010 年 10 月；84(20):10907-12。 Epub 2010 年 8 月 4 日。PubMed PMID：20686015；考研中心。 PMCID：PMC2950557。