TNT: Targeting and Triggering Adjuvancy of DNA Vaccines

TNT：DNA 疫苗的靶向和触发佐剂

• 批准号: 6896249
• 负责人: Michele Aileen Kutzler
• 金额: $ 4.99万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-06-01 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6896249
• 关键词: CD antigens; CD4 molecule; CD44 molecule; CD8 molecule; MHC class I antigen; MHC class II antigen; T lymphocyte; active immunization; antigen antibody reaction; antigen presenting cell; cellular immunity; chemokine; enzyme linked immunosorbent assay; flow cytometry; human immunodeficiency virus 1; immunity; interferon gamma; interleukin 8; laboratory mouse; postdoctoral investigator; tumor necrosis factor alpha; vector vaccine; virus antigen

DESCRIPTION (provided by applicant): To control HIV-1, it is thought that the induction of cell-mediated immunity may be critical for interrupting the establishment of infection. This proposal aims to identify a specific immunization protocol in which vaccination with combinations of DNA constructs encoding the chemokines RANTES, IL-8, or CTACK, and/or a novel B7DeltaC-domain costimulatory protein results in an enhanced antigen specific cell mediated immune response to the model HIV-1 Gag antigens. These novel adjuvants were selected because together they should be able to "target and trigger" (TNT) the immune response in vivo. The goal of this application will be to determine the best method for generating a TNT effect, and secondly, to determine the specific cell phenotypes of the responsible mediators. Several HIV-1 Gag DNA immunization strategies including adjuvant combinations with simultaneous intramuscular injections, sequential prime/boost regime, or alternative immunization timing schedules will be examined. Cellular and humoral immune responses will be measured quantitatively through the use of antibody titer ELISA, IFN-g ELISPOT, and Flow Cytometric staining (intracellular stains for IFN-g and TNF-a, and surface staining for the T cell markers CD45RO, CD27, CCR7, CCR5, and MHC Class I and II Tetramers). Experiments will be carried out in a novel CHAD (transgenic for human MHC Class I and II) murine model system developed in our laboratory. In vivo expression and functional kinetics of antigen and adjuvant in muscle cells will be determined by an immunohistochemical assay, using the unique GeneGrip reporter plasmid encoding GFP. Taken together, this proposal aims to identify and characterize a unique TNT adjuvant combination for HIV-1 DNA vaccines resulting in enhanced antigen specific cell-mediated immune responses and improved HIV-1 DNA vaccine candidates.

描述（由申请人提供）：为了控制HIV-1，细胞介导的免疫的诱导对于中断感染的建立可能是至关重要的。该提案旨在确定一种特定的免疫方案，其中使用编码趋化因子 RANTES、IL-8 或 CTACK 和/或新型 B7DeltaC 结构域共刺激蛋白的 DNA 构建体组合进行疫苗接种，可增强抗原特异性细胞介导的免疫应答HIV-1 Gag 抗原模型。选择这些新型佐剂是因为它们一起应该能够“靶向并触发”（TNT）体内免疫反应。该应用的目标是确定产生 TNT 效应的最佳方法，其次确定负责介质的特定细胞表型。将检查几种 HIV-1 Gag DNA 免疫策略，包括佐剂组合与同时肌内注射、顺序初免/加强方案或替代免疫计时方案。将通过使用抗体滴度 ELISA、IFN-g ELISPOT 和流式细胞术染色（IFN-g 和 TNF-a 的细胞内染色，以及 T 细胞标记 CD45RO、CD27、 CCR7、CCR5 和 MHC I 类和 II 类四聚体）。实验将在我们实验室开发的新型 CHAD（人类 MHC I 类和 II 类转基因）小鼠模型系统中进行。使用编码 GFP 的独特 GeneGrip 报告质粒，通过免疫组织化学测定确定肌肉细胞中抗原和佐剂的体内表达和功能动力学。总而言之，该提案旨在确定和表征 HIV-1 DNA 疫苗的独特 TNT 佐剂组合，从而增强抗原特异性细胞介导的免疫反应并改进 HIV-1 DNA 候选疫苗。

项目成果

Regression of subcutaneous B16 melanoma tumors after intratumoral delivery of an IL-15-expressing plasmid followed by in vivo electroporation.

瘤内递送表达 IL-15 的质粒并随后体内电穿孔后皮下 B16 黑色素瘤肿瘤的消退。

• 发表时间: 2006-10
• 影响因子: 6.4
• 作者: Ugen, K E;Kutzler, M A;Marrero, B;Westover, J;Coppola, D;Weiner, D B;Heller, R
• 通讯作者: Heller, R

Plasmids encoding the mucosal chemokines CCL27 and CCL28 are effective adjuvants in eliciting antigen-specific immunity in vivo.

编码粘膜趋化因子CCL27和CCL28的质粒是在体内引发抗原特异性免疫的有效佐剂。

• 发表时间: 2010-01
• 影响因子: 5.1
• 作者: Kutzler, M A;Kraynyak, K A;Nagle, S J;Parkinson, R M;Zharikova, D;Chattergoon, M;Maguire, H;Muthumani, K;Ugen, K;Weiner, D B
• 通讯作者: Weiner, D B

Systemic immunization with CCL27/CTACK modulates immune responses at mucosal sites in mice and macaques.

使用 CCL27/CTACK 进行全身免疫可调节小鼠和猕猴粘膜部位的免疫反应。

• 发表时间: 2010-02-23
• 影响因子: 5.5
• 作者: Kraynyak, Kimberly A;Kutzler, Michele A;Cisper, Neil J;Khan, Amir S;Draghia;Sardesal, Niranjan Y;Lewis, Mark G;Yan, Jian;Weiner, David B
• 通讯作者: Weiner, David B

Generation of antigen-specific immunity following systemic immunization with DNA vaccine encoding CCL25 chemokine immunoadjuvant.

使用编码 CCL25 趋化因子免疫佐剂的 DNA 疫苗进行全身免疫后产生抗原特异性免疫。

• 发表时间: 2012-11-01
• 影响因子: 4.8
• 作者: Kathuria, Noshin;Kraynyak, Kimberly A;Carnathan, Diane;Betts, Michael;Weiner, David B;Kutzler, Michele A
• 通讯作者: Kutzler, Michele A