ENHANCED T AND B CELL RESPONSES VIA RECOMBINANT PROTEINS

通过重组蛋白增强 T 和 B 细胞反应

• 批准号: 6213323
• 负责人: Edmund J Gosselin
• 金额: $ 23.25万
• 依托单位: ALBANY MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-08-01 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6213323
• 关键词: AIDS vaccines; B lymphocyte; HIV envelope protein gp120; antibody receptor; cytotoxic T lymphocyte; dendritic cells; genetically modified animals; helper T lymphocyte; hepatitis B antigens; human immunodeficiency virus; human subject; interleukin 2; laboratory mouse; leukocyte activation /transformation; macrophage; recombinant proteins; vaccine development

DESCRIPTION: (Adapted from Applicant's Abstract) A safe and successful vaccine against HIV will likely require the simultaneous priming of both cellular and humoral immune responses, and will preferentially involve the use of recombinant proteins. Targeting immunogens to Fc gamma receptor type I (FcgRI) on antigen presenting cells (APC) significantly enhances T cell activation in vitro, and antibody production in vivo. In addition, it can also lead to simultaneous priming of both cytotoxic and helper T cell responses. Furthermore, by combining the administration of antigen with cytokines, T cell activation can be further enhanced, and T cell subset development modulated. It has also been demonstrated that targeting antigen (Ag) to FcgRI on APC can eliminate the need for traditional adjuvant, easing difficulties associated with vaccine preparation and distribution. Therefore, developing a strategy which facilitates antigen targeting to APC, and the use of cytokines in vaccines, is likely to have a significant impact on current vaccine technology, in particular as it applies to HIV. We propose to utilize molecular techniques, and FcgRI-specific constructs, to create and test the ability of a prototype two component (modular) immune targeting system to stimulate enhanced humoral, CD4 helper T cell, and CD8 cytotoxic T cell responses in vitro and in vivo. Components will consist of a humanized divalent FcgRI-specific biotin-binding targeting element, and biotinylated functional elements including Hepatitis B Ag, gp120 Ag, and IL-2. The ability of the two component immunogens to modulate human CD4 and CD8 T cell responses in vitro, and murine B cell, CD4 T cell, and CD8 T cell responses in vivo, will be examined. In the latter instance, transgenic mice that express human FcgRI will be immunized with two component immunogens. Following immunization, CD4 and CD8 T cell responses, as well as the generation of Ag-specific antibody will be measured. These studies will provide a novel and safe approach for simultaneously priming humoral and cellular responses in vivo using recombinant proteins. This approach will not only provide an effective means for controlling the spread of HIV, but many other infectious organisms as well.

描述：(改编自申请者摘要)一种安全、成功的疫苗 抗艾滋病毒很可能需要同时启动细胞和 体液免疫反应，并将优先涉及使用 重组蛋白。免疫原靶向Fc-γ受体I型(FcgRI) 抗原提呈细胞(APC)显著增强T细胞活化 体外，体内产生抗体。此外，它还可能导致 同时启动细胞毒性和辅助T细胞反应。 此外，通过结合给予抗原和细胞因子，T细胞 可以进一步增强激活，调节T细胞亚群的发育。它 APC上针对FcgRI的靶向抗原(Ag)也被证明可以 消除了对传统佐剂的需求，缓解了相关困难 疫苗的准备和分发。因此，制定一项战略 这有助于抗原靶向APC，并在 疫苗，可能会对目前的疫苗技术产生重大影响， 特别是当它适用于艾滋病毒时。我们建议利用分子技术， 和FcgRI特定的构造，以创建和测试原型的能力 双组分(模块化)免疫靶向系统，以刺激增强的体液， 体外和体内的CD4辅助T细胞和CD8细胞毒性T细胞反应。 组件将由人源化的二价FcgRI-特异性生物素结合组成 靶向元件和包括乙肝在内的生物素化功能元件 Ag、gp120 Ag和IL-2。两种组分免疫原的调节能力 人CD4和CD8T细胞体外反应，小鼠B细胞，CD4T细胞，和 体内的CD8 T细胞反应，将被检测。在后一种情况下， 表达人FcgRI的转基因小鼠将用两种成分免疫 免疫原。免疫后，CD4和CD8 T细胞反应以及 将测量抗原特异性抗体的生成。这些研究将 提供了一种新颖而安全的方法，同时启动体液和 使用重组蛋白的体内细胞反应。这种方法不会 只是提供了控制艾滋病毒传播的有效手段，但许多 其他感染性生物也是如此。