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: 6374417
• 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 2004-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6374417
• 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：（改编自申请人的摘要）一种安全且成功的疫苗 对抗艾滋病毒可能需要同时启动细胞和 体液免疫反应，并优先涉及使用 重组蛋白。将免疫原靶向 I 型 Fc γ 受体 (FcgRI) 抗原呈递细胞 (APC) 显着增强 T 细胞活化 体外和体内抗体生产。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，通过结合施用抗原和细胞因子，T细胞 激活可以进一步增强，并且 T 细胞亚群的发育受到调节。它 还已证明，将抗原 (Ag) 靶向 APC 上的 FcgRI 可以 消除对传统佐剂的需求，缓解相关困难 疫苗的制备和分发。因此，制定战略 这有助于抗原靶向 APC，以及细胞因子在 疫苗，可能会对当前的疫苗技术产生重大影响， 特别是因为它适用于艾滋病毒。我们建议利用分子技术， 和 FcgRI 特定构建体，以创建和测试原型的能力 两部分（模块化）免疫靶向系统刺激增强的体液， CD4 辅助 T 细胞和 CD8 细胞毒性 T 细胞在体外和体内的反应。 组件将由人源化二价 FcgRI 特异性生物素结合组成 靶向元件和生物素化功能元件，包括乙型肝炎 Ag、gp120 Ag 和 IL-2。两种成分免疫原的调节能力 体外人类 CD4 和 CD8 T 细胞反应，以及鼠 B 细胞、CD4 T 细胞和 将检查体内 CD8 T 细胞反应。在后一种情况下， 表达人 FcgRI 的转基因小鼠将用两种成分进行免疫 免疫原。免疫后，CD4 和 CD8 T 细胞反应以及 将测量Ag特异性抗体的产生。这些研究将 提供一种新颖且安全的方法来同时启动体液和 使用重组蛋白进行体内细胞反应。这种方法不会 仅仅提供了控制艾滋病毒传播的有效手段，但许多 其他传染性生物体也是如此。