RNAi manipulations of DC to enhance HIV immunogenicity

DC 的 RNAi 操作增强 HIV 免疫原性

基本信息

批准号：
7761032
负责人：
Premlata Shankar
金额：
$ 38.58万
依托单位：
TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-15 至 2014-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7761032
关键词：
Adjuvant Alleles Animal Model Antibodies Antigen-Presenting Cells Antigens Binding CD8B1 gene Cells Clinical Trials Dendritic Cells Development Drug Formulations Engineering Epidemic Epitopes Failure Gagging Gene Targeting Generations Genes HIV HIV Antigens HIV Infections HIV vaccine HLA-A2 Antigen HLA-B27 Antigen Human Immune Immune response Immunity Immunization Immunodeficient Mouse Immunologic Adjuvants In Vitro Infection Integrins Interleukin 2 Receptor Interleukin 2 Receptor Gamma Interleukin-10 Intervention Leukocytes Ligands Liposomes Mediating Messenger RNA Methods Modeling Molecular Monkeys Mouse Strains Mus Mutation Pan Genus Peptide antibodies Peptides Peripheral Physiological Positioning Attribute Pre-Clinical Model Proteins Publishing RNA Interference Reagent Recombinants Small Interfering RNA T-Lymphocyte Testing Therapeutic Transgenic Organisms Treatment Protocols Vaccines Viral Viral Vector Virulent Virus animal tissue base cytokine design efficacy testing immunogenicity improved in vivo inhibitor/antagonist insight melanoma mouse model novel novel strategies novel vaccines pre-clinical prevent response targeted delivery tool tool development vector-based vaccine

项目摘要

DESCRIPTION (provided by applicant): To date there is no effective vaccine for HIV infection. Although a major vaccine trial based on homologous recombinant viral prime/boost failed recently, a study undertaken later, in a monkey model, suggests that with a more appropriate heterologous prime/boost regimen, it is possible to elicit a strong T cell response that protects against virulent viral challenge. Thus, for a vaccine to be effective, it should be able to evoke a strong and broad-based T cell response. Moreover, pretesting the relevance of novel vaccine approaches by HIV challenge in preclinical models would greatly help prevent the agony of vaccine failures in human clinical trials. As Dendritic cells are critical for induction of T cell immune responses, we hypothesize that immunization with HIV proteins targeted to dendritic cells in which select negative immunomodulatory molecules such as SOCS-1, PD-L1, L2 and IL-10 have been suppressed by RNA interference will elicit a potent polyfunctional CD8+ T cell response. Our hypothesis is based on our preliminary results in which silencing of SOCS-1 via targeted siRNA delivery to DC was enough to elicit a robust primary T cell response in vitro to several HIV gag epitopes, including subdominant ones. Moreover, we have recently shown the feasibility of using the latest versions of humanized mouse models to test the efficacy of siRNA mediated interventions in HIV infection and are thus are in a position to test whether the human DC- targeted methods are effective in vivo. In Specific Aim 1 of this proposal we will develop methods and reagents for targeted delivery of HIV-antigens and immunomodulatory siRNA reagents to human DCs. These will include a DC targeting peptide modified to bind siRNA as well as to deliver HIV antigens, two DC-targeting antibody fused to HIV proteins and further modified to bind siRNAs and a liposomal formulation that allows targeted delivery of siRNA and HIV antigen in mRNA form. In Aim 2, we will evaluate whether co-delivery of HIV immunogen with the different immunomodulatory siRNA (singly and in combination) by any of these methods is able to induce a broad and polyfunctional primary HIV-specific CD8 T cell response in vitro. In Aim 3, we will validate the in vitro findings as well as test the efficacy of our methods to actually confer protection from in vivo HIV challenge in the humanized BLT mouse model transgenic for HLA-A2 and HLA-B27. This project will develop DC-targeted approaches for HIV antigen delivery and RNAi manipulations as a novel HIV vaccine strategy to induce potent virus-specific T cell immunity. If successful, the studies would provide an alternative to the recombinant viral vector-based vaccines. In addition, the in vivo studies in a preclinical animal model will provide novel insights into the immune correlates of protection.

描述（由申请人提供）：迄今为止，没有有效的艾滋病毒感染疫苗。尽管最近一项基于同源重组病毒素/增强的主要疫苗试验失败了，但后来在猴子模型中进行的一项研究表明，使用更合适的异源质量/增强方案，可以引起强大的T细胞反应，以防止病毒攻击。因此，为了使疫苗有效，它应该能够引起强大且广泛的T细胞反应。此外，在临床前模型中，通过HIV挑战的新型疫苗方法的测试相关性将极大地帮助防止人类临床试验中的疫苗失败的痛苦。由于树突状细胞对于诱导T细胞免疫反应至关重要，因此我们假设用针对树突状细胞的HIV蛋白进行免疫，其中选择的阴性免疫调节分子（例如SOCS-1，PD-L1，L2，L2和IL-10）被RNA抑制了RNA抑制，RNA抑制了有效的cd8+ T细胞响应。我们的假设是基于我们的初步结果，在该结果中，SOCS-1通过靶向siRNA向DC进行沉默足以在体外引起对几种HIV GAG表位（包括亚辅助）的强大原代T细胞反应。此外，我们最近显示了使用最新版本的人源性小鼠模型测试siRNA介导的HIV感染中的疗效，因此可以测试人类DC靶向方法是否在体内有效。在该提案的特定目的1中，我们将开发针对人类DC靶向递送HIV-抗原和免疫调节siRNA试剂的方法和试剂。这些将包括一个靶向肽修饰以结合siRNA的DC靶向肽以及递送HIV抗原，两种融合到HIV蛋白的DC靶向抗体，并进一步修饰以结合siRNAS和脂质体配方，允许以mRNA形式靶向靶向siRNA和HIV抗原。在AIM 2中，我们将评估与不同免疫调节性siRNA（单独和组合）通过任何这些方法共同传递HIV免疫原能否能够在体外诱导广泛的多功能原发性HIV特异性CD8 T细胞反应。在AIM 3中，我们将验证体外发现，并测试我们方法在HLA-A2和HLA-B27的人源化BLT小鼠模型转基因中实际赋予体内HIV挑战的疗效。该项目将开发针对DC的HIV抗原递送和RNAi操纵的方法，作为一种新型的HIV疫苗策略，以诱导有效的病毒特异性T细胞免疫。如果成功，这些研究将为基于重组病毒载体的疫苗提供替代方案。此外，临床前动物模型中的体内研究将提供有关保护的免疫相关性的新见解。