Integrase-defective lentiviral based influenza vaccines

整合酶缺陷型慢病毒流感疫苗

• 批准号: 7895034
• 负责人: MIRELLA SALVATORE
• 金额: $ 22.24万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-18 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7895034
• 关键词: Address; Adjuvant; Animal Model; Animals; Antigen Targeting; Antigens; Body Weight decreased; CD8B1 gene; Cell Culture Techniques; Cell Nucleus; Cells; Cellular Immunity; Characteristics; Circular DNA; Cytotoxic T-Lymphocytes; DNA; DNA Integration; DNA Vaccines; DNA delivery; Data; Dendritic Cells; Effectiveness; Exhibits; Gene Transfer; Generations; Genes; Genetic Materials; Genome; Helper-Inducer T-Lymphocyte; Human; Immune response; Immune system; Immunity; Immunization; Immunologics; Infection; Influenza; Influenza A virus; Integrase; Interphase Cell; Intramuscular Injections; Kinetics; Lentivirus Vector; Lung; Mediating; Membrane Proteins; Methods; Modeling; Morbidity - disease rate; Mus; Mutation; NP protein; Nucleoproteins; Plasmids; Promoter Regions; Proteins; Recombinants; Reporter; Risk; Role; Safety; System; Testing; Time; Transcriptional Regulation; Vaccination; Vaccines; Viral; Viral Vector; Viral load measurement; Virus Diseases; base; cell mediated immune response; cell type; cellular transduction; efficacy testing; extrachromosomal DNA; gene gun; improved; in vivo; influenza A virus nucleoprotein; influenza virus vaccine; influenzavirus; interest; mouse model; novel; protective efficacy; protein expression; public health relevance; research study; response; vaccine development; vector; vector-induced; vesicular stomatitis virus G protein

DESCRIPTION (provided by applicant): A key problem in vaccine development is finding an effective way to induce cellular immunity. DNA vaccines are constituted by a plasmid encoding the target antigen under the transcriptional control of a promoter region active in human cells. They are good inducers of cellular immune responses however they require a high amount of DNA and three administrations to be effective. Lentiviral vectors (IC-LV) are powerful inducers of cell mediated and humoral immune responses, however they carry the intrinsic risk associated with integration into the host genome. Non-integrating lentiviral vectors (ID-LV) have a mutation in the integrase gene, do not integrate into the genome, nor do they replicate, but are maintained as episomal circular DNA in the nucleus (E-DNA) in the absence of integration. We have previously shown that E-DNA is stable and can express functional proteins. Proteins expressed by ID-LV are recognized by the immune system in cell culture. In addition, ID-LV expressing GFP elicited a long-lasting immune response to GFP after a single intramuscular injection in vivo suggesting that a single administration may be sufficient to induce a protective response. As proof-of-concept of the effectiveness of ID-LV, we will compare the responses induced by ID-LV expressing the NP protein of influenza virus with those obtained with conventional DNA vaccines and IC-LV. Use of influenza virus offers several advantages. First, cell-mediated immunity confers protection from multiple strains towards viral challenge. Second, DNA vaccines have been proven to be effective in inducing protective cell-mediated immunity in animal models. In Aim 1 we will compare humoral and cellular immune response, elicited by immunization with ID-LV, IC-LV and DNA vaccine expressing influenza NP. In Aim 2 we will test the ability of ID-LV to protect the animal from sublethal or lethal challenge and address the question of cross-protection. The influenza mouse model provides the first opportunity to test the efficacy of a protein expressed from ID-LV to protect from a viral challenge in vivo and to compare both protection efficacy and immunologic parameters among the non-integrated and integrated vector and a conventional DNA vaccine. These studies will define the general utility of ID-LV as a delivery vector. If successful, this approach would represent a significant step in vaccine development given the safety feature of non integration. PUBLIC HEALTH RELEVANCE: We have previously shown that proteins expressed by integrase-defective lentiviral vectors (ID-LV) are recognized by the immune system in cell culture and can elicit a long-lasting immune response after a single intramuscular injection in vivo. In this proposal, as proof-of-concept of the effectiveness of ID-LV as a vaccine, we will compare the responses induced by ID-LV expressing the NP protein of influenza virus with those obtained with conventional DNA vaccines and integrase-competent-LV. The influenza mouse model provides the first opportunity to test the efficacy of a protein expressed from ID-LV to protect from a viral challenge in vivo and to compare both protection efficacy and immunologic parameters among the ID-LV and IC-LV and a conventional DNA vaccine. These studies will define the utility of ID-LV as a delivery vector. If successful, this approach would represent a significant step in vaccine development given the safety feature of non-integration.

描述（由申请人提供）：疫苗开发中的一个关键问题是找到诱导细胞免疫的有效方法。DNA疫苗由编码靶抗原的质粒构成，所述靶抗原在人细胞中有活性的启动子区的转录控制下。它们是细胞免疫应答的良好诱导剂，然而它们需要大量的DNA和三次施用才有效。慢病毒载体（IC-LV）是细胞介导和体液免疫应答的强有力诱导剂，然而它们携带与整合到宿主基因组中相关的内在风险。非整合型慢病毒载体（ID-LV）在整合酶基因中具有突变，不整合到基因组中，也不复制，但在不整合的情况下作为附加型环状DNA（E-DNA）维持在细胞核中。我们以前已经证明E-DNA是稳定的，可以表达功能蛋白。由ID-LV表达的蛋白质在细胞培养物中被免疫系统识别。此外，表达GFP的ID-LV在体内单次肌内注射后引发对GFP的持久免疫应答，表明单次施用可能足以诱导保护性应答。作为ID-LV有效性的概念验证，我们将比较表达流感病毒NP蛋白的ID-LV诱导的应答与常规DNA疫苗和IC-LV诱导的应答。使用流感病毒有几个优点。首先，细胞介导的免疫赋予针对病毒攻击的多种毒株的保护。其次，DNA疫苗已被证明在动物模型中有效诱导保护性细胞介导的免疫。在目的1中，我们将比较由ID-LV、IC-LV和表达流感NP的DNA疫苗免疫引起的体液和细胞免疫应答。在目标2中，我们将测试ID-LV保护动物免受亚致死或致死攻击的能力，并解决交叉保护问题。流感小鼠模型提供了第一次机会来测试从ID-LV表达的蛋白质在体内保护免受病毒攻击的功效，并比较非整合的和整合的载体与常规DNA疫苗之间的保护功效和免疫学参数。这些研究将确定ID-LV作为递送载体的一般效用。如果成功，这种方法将代表疫苗开发的重要一步，因为它具有非整合的安全性。 公共卫生关系：我们以前已经表明，由整合酶缺陷型慢病毒载体（ID-LV）表达的蛋白质在细胞培养中被免疫系统识别，并且在体内单次肌内注射后可以引起持久的免疫应答。在本提案中，作为ID-LV作为疫苗的有效性的概念验证，我们将比较表达流感病毒NP蛋白的ID-LV诱导的应答与用常规DNA疫苗和整合酶活性LV获得的应答。流感小鼠模型提供了第一次机会来测试从ID-LV表达的蛋白质在体内保护免受病毒攻击的功效，并比较ID-LV和IC-LV与常规DNA疫苗之间的保护功效和免疫学参数。这些研究将确定ID-LV作为递送载体的效用。如果成功，这种方法将代表疫苗开发的重要一步，因为它具有非整合的安全性。