Development of a DNA Vaccine against RSV

针对 RSV 的 DNA 疫苗的开发

• 批准号: 7113678
• 负责人: SHREE R SINGH
• 金额: $ 9.55万
• 依托单位: ALABAMA STATE UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7113678
• 关键词: CHO cells; SDS polyacrylamide gel electrophoresis; active immunization; biotechnology; cellular immunity; enzyme linked immunosorbent assay; flow cytometry; fluorescence microscopy; humoral immunity; immune response; laboratory mouse; molecular cloning; mucosal immunity; respiratory disorder chemotherapy; respiratory syncytial virus; tissue /cell culture; vaccine development; vaccine evaluation; vector vaccine; viral vaccines; virus load; western blottings

The Proposed project goal is to develop and study a DNA vaccine against Respiratory Syncytial Virus (RSV). RSV infects almost all children under two years of age and there is no vaccine available. This project aims to engineer a recombinant DNA vector containing immunologically important regions of the RSV F gene and modified cholera toxin gene. It is expected that the DNA immunogen will induce local and systemic immune response when administered intramuscularly. Since DNA plasmids induce a Th1 cell response, this approach is particularly well suited against RSV where Th1 immune response is very critical in inducing protective immunity. Attractive aspects of the project include the simplicity of the design and development of the DNA plasmid, potential to induce cellular, humoral and the mucosal immune responses. The DNA plasmid will be constructed using a DNA vector (phCMV1 from Gene Therapy Systems) into which regions of RSV F and modified cholera toxin genes will be cloned. The phCMV1 DNA vector contains a CMV promoter, intron A and a strong terminator. DNA will be purified and delivered in mice intramuscularly. Additionally, to test the establishment of Th1 cell response, mice will be immunized with combination of DNA vaccine and recombinant RSV F protein. The humoral and cellular immune responses resulting from immunizations will be analyzed. The ability of the immunogen to protect mice from RSV infection will be evaluated by challenging mice with live RSV. The upper and lower respiratory tract infections will be assessed by studying virus load in upper mucosa and the lungs. This study will lead to an understanding of the DNA vaccination and mucosal immune response and contribute to design and development of a safer and more effective vaccine against RSV and other viruses infecting the mucosa.

该项目的目标是开发和研究针对呼吸道合胞病毒（RSV）的DNA疫苗。RSV感染几乎所有2岁以下的儿童，并且没有可用的疫苗。本项目旨在设计一种含有RSV F基因的免疫学重要区域和经修饰的霍乱毒素基因的重组DNA载体。预期DNA免疫原在肌内给药时将诱导局部和全身免疫应答。由于DNA质粒诱导Th1细胞应答，该方法特别适合于抗RSV，其中Th1免疫应答在诱导保护性免疫中非常关键。该项目吸引人的方面包括DNA质粒的设计和开发的简单性，诱导细胞、体液和粘膜免疫应答的潜力。DNA质粒将使用DNA载体（来自Gene Therapy Systems的phCMV 1）构建，RSV F和修饰的霍乱毒素基因的区域将被克隆到该DNA载体中。phCMV 1 DNA载体含有CMV启动子、内含子A和强终止子。DNA将被纯化并输送到小鼠体内 肌肉注射此外，为了测试Thl细胞应答的建立，将用DNA疫苗和重组RSV F蛋白的组合免疫小鼠。将分析免疫接种产生的体液和细胞免疫应答。将通过用活RSV攻击小鼠来评价免疫原保护小鼠免受RSV感染的能力。将通过研究上粘膜和肺中的病毒载量来评估上呼吸道和下呼吸道感染。这项研究将导致对DNA疫苗接种和粘膜免疫应答的理解，并有助于设计和开发一种更安全，更有效的疫苗，以对抗RSV和其他感染粘膜的病毒。