Protective Activity of a Multi-Functional Immunogen

需要目前人类佐剂的替代品。我们的研究表明，在缺乏 抗原呈递细胞上人Fc γ受体I型（hFc γ RI）的靶向抗原（Ag）佐剂 增强体外T细胞活化和体内Ag特异性抗体（Ab）和细胞因子的产生。然而，在这方面， 还没有人创造出重组免疫原，其允许检测hFc γ RI靶向 在传染病模型中的方法，或所涉及的机制的故意研究。链球菌- 肺炎球菌是一种呼吸道病原体，已鉴定出其保护性Ag（PspA）。PspA是 一种细菌表面蛋白，当与佐剂一起用作免疫原时， S.肺炎感染小鼠。我们假设：1）保护S。肺炎感染可以是 通过用抗hFc γ RI-PspA融合蛋白免疫增强（在没有佐剂的情况下）。（二） 增强的机制也将涉及Ag在淋巴组织中定位的改变， 作为银加工的改变。为了测试后者，我们将产生抗hFc γ RI-HEL融合蛋白。 HEL是一种蛋白质Ag，通常用于进行Ag加工和呈递的机制研究， 体外和体内。A）将抗hFc γ RI-PspA和抗hFc γ RI-PspA去人源化， HEL融合蛋白已经产生。B）检查融合蛋白增强T和B细胞增殖的能力。 在体外和体内的反应。C）在体内确定hFc γ RI靶向Ag是否部分地通过以下方式起作用： 增强Ag对淋巴组织的定位。在具体目标2中，我们将：A）测试抗- hFc γ RI-PspA融合蛋白保护免受S.肺炎。B）研究影响 hFc γ RI靶向Ag加工和呈递，利用生物化学、免疫学和免疫学方法， 超微结构技术这些研究将为进一步靶向hFc γ RI奠定基础 在小鼠和人类中使用各种传染病病原体的研究，消除了对传统方法的需要， 佐剂，实质上减少接种个体所需的Ag的量（降低成本和 潜在的毒性），并提供一种疫苗递送系统，其同时增强体液和 细胞免疫反应。因此，这些研究和这种疫苗策略将适用于疫苗 在成人、儿童和免疫功能低下人群中对抗多种病原体。

Alternatives to current human adjuvants are needed. Our studies demonstrate that in the absence of adjuvant, targeting antigen (Ag) to human Fcgamma receptor type I (hFcgammaRI) on Ag presenting cells enhances T cell activation in vitro and Ag-specific antibody (Ab) and cytokine production in vivo. However, no one has created recombinant immunogens, which permit examination of the hFcgammaRI targeting approach in an infectious disease model, or the deliberate study of the mechanism(s) involved. Strepto- coccus pneumoniae is a respiratory pathogen for which a protective Ag (PspA) has been identified. PspA is a bacterial surface protein, which, when used as an immunogen with adjuvant, generates protection against S. pneumoniae infection in mice. We hypothesize: 1) That protection against S. pneumoniae infection can be enhanced (in the absence of adjuvant) by immunizing with an anti-hFcgammaRI-PspA fusion protein. 2) That the mechanism(s) of enhancement will involve alterations in Ag localization to lymphoid tissues, as well as alterations in Ag processing. To test the latter, we will generate an anti-hFcgammaRI-HEL fusion protein. HEL is a protein Ag commonly used to conduct mechanistic studies of Ag processing and presentation, in vitro and in vivo. In Speific Aim 1 we will: A) Dehumanize anti-hFcgammaRI-PspA and anti-hFcgammaRI- HEL fusion proteins already generated. B) Examine the ability of the fusion proteins to enhance T and B cell responses in vitro and in vivo. C) Determine, in vivo, if hFcgammaRI targeting of Ag works, in part, through enhanced localization of Ag to lymphoid tissues. In Specific Aim 2 we will: A) Test the ability of the anti- hFcgammaRI-PspA fusion protein to protect against challenge with S. pneumoniae. B) Study the influence of hFcgammaRI targeting on Ag processing and presentation utilizing biochemical, immunological and ultrastructural techniques. The proposed studies will lay the ground-work for further hFcgammaRI targeting studies using a variety of infectious disease agents in mouse and human, eliminate the need for traditional adjuvant, substantially reduce the amount of Ag required to vaccinate an individual (reducing cost and potential toxicity), and provide a vaccine delivery system, which simultaneously enhances humoral and cellular immune responses. Thus, these studies and this vaccine strategy will be applicable to vaccines against a wide variety of pathogens in adult, pediatric, and immunocompromised populations.