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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：
8317541
负责人：
Premlata Shankar
金额：
$ 36.64万
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/8317541
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 Gamma Interleukin-10 Interleukin-2 Intervention Leukocytes Ligands Liposomes Mediating Messenger RNA Methods Modeling Molecular Monkeys Mouse Strains Mus Mutation Peptide antibodies Peptides Peripheral Physiological Positioning Attribute Pre-Clinical Model Proteins Publishing RNA Interference Reagent Recombinants Regimen Small Interfering RNA T cell response T-Lymphocyte Testing Therapeutic Transgenic Organisms Vaccines Viral 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 receptor recombinant viral vector 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.

说明（由申请人提供）：迄今为止，没有有效的艾滋病毒感染疫苗。尽管一项基于同源重组病毒引物/增强的主要疫苗试验最近失败了，但后来在猴子模型中进行的一项研究表明，采用更合适的异源引物/增强方案，有可能引起强烈的T细胞反应，从而保护人体免受毒力病毒的攻击。因此，为了使疫苗有效，它应该能够引起强烈和广泛的T细胞反应。此外，在临床前模型中通过HIV挑战预先测试新疫苗方法的相关性将大大有助于防止疫苗在人体临床试验中失败的痛苦。由于树突状细胞对诱导T细胞免疫应答至关重要，我们假设HIV蛋白免疫靶向树突状细胞，其中选择了被RNA干扰抑制的负免疫调节分子，如SOCS-1、PD-L1、L2和IL-10，将引发有效的多功能CD8+ T细胞应答。我们的假设是基于我们的初步结果，即通过靶向siRNA递送到DC来沉默SOCS-1足以在体外引发对几种HIV gag表位（包括亚显性表位）的强大原代T细胞反应。此外，我们最近已经证明了使用最新版本的人源化小鼠模型来测试siRNA介导的干预HIV感染的有效性的可行性，因此能够测试人类DC靶向方法是否在体内有效。在本提案的Specific Aim 1中，我们将开发hiv抗原和免疫调节siRNA试剂靶向递送到人类dc的方法和试剂。这些将包括一种经过修饰的DC靶向肽，可以结合siRNA并递送HIV抗原，两种DC靶向抗体融合到HIV蛋白并进一步修饰以结合siRNA，以及一种脂质体制剂，允许以mRNA形式靶向递送siRNA和HIV抗原。在Aim 2中，我们将评估通过这些方法中的任何一种，将HIV免疫原与不同的免疫调节siRNA（单独或联合）共同递送是否能够在体外诱导广泛和多功能的原发性HIV特异性CD8 T细胞反应。在目标3中，我们将验证体外研究结果，并测试我们的方法在转基因HLA-A2和HLA-B27的人源化BLT小鼠模型中对体内HIV攻击提供保护的有效性。