ENHANCED T AND B CELL RESPONSES VIA RECOMBINANT PROTEINS

描述：（改编自申请人摘要）一种安全和成功的疫苗 对抗艾滋病毒可能需要同时启动细胞和 体液免疫反应，并将优先涉及使用 重组蛋白将免疫原靶向至I型Fc γ受体（FcgRI） 抗原提呈细胞（APC）上的T细胞活化显着增强， 体外和体内抗体产生。此外，它还可能导致 同时引发细胞毒性和辅助性T细胞应答。 此外，通过将抗原与细胞因子联合施用，T细胞 活化可以进一步增强，并且调节T细胞亚群发育。它 也已经证明，将抗原（Ag）靶向APC上的FcgRI， 消除了对传统佐剂的需要，缓解了相关困难， 疫苗的制备和分发。因此，制定战略 其促进抗原靶向APC，以及细胞因子在 疫苗，可能会对目前的疫苗技术产生重大影响， 特别是在艾滋病毒方面。我们建议利用分子技术， 和FcgRI特异性构建体，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， CD 4辅助性T细胞和CD 8细胞毒性T细胞应答。 组分将由人源化二价FcgRI特异性生物素结合多肽组成。 靶向元件和生物素化的功能元件，包括B型肝炎 Ag、gp 120 Ag和IL-2。两种成分免疫原调节 体外人CD 4和CD 8 T细胞应答，以及鼠B细胞、CD 4 T细胞和 将检查体内CD 8 T细胞应答。在后一种情况下， 表达人FcgRI转基因小鼠将用两种组分免疫 免疫原免疫后，CD 4和CD 8 T细胞应答，以及 测定Ag特异性抗体的产生。这些研究将 提供了一种新的安全的方法， 使用重组蛋白的体内细胞应答。这种做法不会 这只是控制艾滋病毒传播的有效手段，但许多 还有其他传染性微生物

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.