DC-targeted DNA vaccines

DC 靶向 DNA 疫苗

• 批准号: 7774820
• 负责人: Ralph Marvin Steinman
• 金额: $ 43.51万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-25 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7774820
• 关键词: AIDS Vaccines; Adjuvant; Antibodies; Antibody Formation; Antigen Presentation; Antigen Targeting; Antigens; Area; B-Lymphocytes; Biological Assay; CD4 Positive T Lymphocytes; Cell Differentiation process; Cell Maturation; Cell physiology; Cells; Cellular Immunity; Cellular biology; Chimeric Proteins; Communities; Core Facility; Cross Presentation; DEC-205 receptor; DNA; DNA Vaccines; Data; Dendritic Cells; Dose; Engineering; Ensure; Fc Receptor; Frequencies; Gagging; Goals; Grant; HIV; HIV vaccine; Haplotypes; Hen Egg Lysozyme; Human; Hybridomas; Immune; Immune response; Immunity; In Situ; Learning; Ligands; Ligation; Light; Lymphoid Tissue; MHC Class I Genes; MHC Class II Genes; Macaca; Macaca mulatta; Mass Spectrum Analysis; Mediating; Memory; Methods; Modeling; Monitor; Monkeys; Monoclonal Antibodies; Mus; Ovalbumin; Pathway interactions; Peptide/MHC Complex; Peptides; Peripheral Blood Mononuclear Cell; Poly I-C; Primates; Process; Production; Property; Proteins; Receptor Cell; Relative (related person); Research; Route; SIV; Signal Transduction; Stimulus; System; T-Lymphocyte; Testing; Transgenic Mice; Transgenic Organisms; Vaccination; Vaccine Antigen; Vaccine Design; Vaccine Research; Vaccines; Work; antigen processing; base; cell mediated immune response; cytokine; design; immunogenicity; improved; in vivo; langerin; monocyte; mouse model; novel strategies; programs; protective efficacy; receptor; research study; response; targeted delivery; trafficking; uptake; vaccine efficacy

We have begun a new approach to vaccines. We directly harness the properties of dendrific cells (DCs) to improve vaccine efficacy, with emphasis on T cell mediated immunity. Previously we designed protein vaccines to improve delivery of HIV gag to DCs in mice. Directed or targeted delivery of gag within a monoclonal antibodies to the DEC-205 receptor on DCs gave a large increase in immunogenicity relative to nontargeted protein as well as protection in a mouse model. A newer example of DC targeting involves DNA vaccines, as will be studied here. The vaccine encodes a fusion protein comprised of a single chain Fv antibody to DCs fused to HIV gag. Immunizing and protective efficacy increased ~100 fold. We have 4 aims to study mechanisms and improve DNA vaccines along with several key collaborators: 1) Delivery of DNA vaccine anfigens to DCs is currently monitored indirectly, by virtue ofthe immunity that is induced. To gain direct data on DC uptake and processing of vaccines in mice, we will a) tag vaccine proteins with a sensitive system we developed called "OLLAS"; b) use antibodies to MHC-peptide, and c) isolate MHC products from targeted DCs to identify the presented pepfides. 2) With these assays we will then compare different DC receptors for targeting vaccine protein to these critical presenting cells. We have prepared hybridomas and cloned the heavy and light chains to additional DC receptors like Langerin/CD207 and DC-SIGN/CD209, so that we can prepare single chain Fv gag fusion DNA vaccines to other DC targets. 3) To improve DNA vaccine efficacy at the level of DCs, we will study the immunostimulatory effects of different DC maturation stimuli, including Ig C region sequences that preferenfially ligate activating FcyR. 4) To extend DC-based DNA vaccines to the SIV macaque model, we already have prepared anti-human DEC-205 antibodies that react with monkey DEC-205 and mediate efficient cross presentation of gag on MHC class I, and we have prepared human DEC-205 transgenic mice to ensure that anti-rhesus/human single chain DEC successfully targets and presents SIV antigens in vivo. This will allow us to determine if single chain Fv gag fusion DNA vaccines successfully target to DCs in monkey lymphoid tissues.

我们已经开始采用新的疫苗方法。我们直接利用树突细胞（DC）的特性来 提高疫苗功效，重点是 T 细胞介导的免疫。之前我们设计了蛋白质 疫苗可改善 HIV gag 向小鼠 DC 的输送。在一定范围内定向或有针对性地进行堵嘴 相对于 DC 上的 DEC-205 受体的单克隆抗体，其免疫原性大大增加 非靶向蛋白质以及小鼠模型中的保护作用。 DC 靶向的一个新例子涉及 DNA 疫苗，正如这里将要研究的那样。该疫苗编码由单链Fv组成的融合蛋白 与 HIV gag 融合的 DC 抗体。免疫和保护功效提高约 100 倍。我们有4个目标 与几个主要合作者一起研究机制并改进 DNA 疫苗：1) DNA 的传递 目前，DC的疫苗抗原是通过诱导的免疫来间接监测的。为了获得 关于小鼠 DC 摄取和疫苗加工的直接数据，我们将 a) 用敏感的标签标记疫苗蛋白 我们开发的系统称为“OLLAS”； b) 使用 MHC 肽抗体，并且 c) 从其中分离 MHC 产物 靶向 DC 来识别所呈现的肽。 2) 通过这些测定，我们将比较不同的 DC 将疫苗蛋白靶向这些关键呈递细胞的受体。我们制备了杂交瘤和 将重链和轻链克隆到其他 DC 受体，如 Langerin/CD207 和 DC-SIGN/CD209，因此 我们可以制备针对其他 DC 靶点的单链 Fv gag 融合 DNA 疫苗。 3) 改善DNA DC水平上的疫苗功效，我们将研究不同DC成熟度的免疫刺激作用 刺激，包括优先连接激活FcγR的Ig C区序列。 4）基于DC扩展 针对SIV猕猴模型的DNA疫苗，我们已经制备了抗人DEC-205抗体， 与猴子 DEC-205 反应并介导 MHC I 类上堵嘴的有效交叉呈递，我们有 制备人DEC-205转基因小鼠，确保抗恒河猴/人单链DEC成功 体内靶向并呈递 SIV 抗原。这将使我们能够确定单链 Fv gag 融合 DNA 疫苗成功地靶向猴淋巴组织中的 DC。