Protective Activity of a Multi-Functional Immunogen

多功能免疫原的保护活性

• 批准号: 7350212
• 负责人: Edmund J Gosselin
• 金额: $ 22.45万
• 依托单位: ALBANY MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-02-15 至 2010-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7350212
• 关键词: Adjuvant; Adult; Antibodies; Antigen Targeting; Antigens; Autologous; B-Lymphocytes; Binding; Biochemical; Biological Assay; Cell Fractionation; Cell Proliferation; Cell physiology; Cell surface; Cells; Childhood; Chimeric Proteins; Cholera Toxin; Communicable Diseases; Complex; Cytometry; DNA; Enzyme-Linked Immunosorbent Assay; Figs - dietary; Flow Cytometry; Generations; Harvest; Hen Egg Lysozyme; Hour; Human; Hybridomas; Immune response; Immunization; Immunocompromised Host; Immunofluorescence Immunologic; Immunohistochemistry; Immunologic Techniques; In Vitro; Individual; Infection; Injection of therapeutic agent; Label; Laboratories; Lethal Dose 50; Lymphoid; Lymphoid Tissue; Measurement; Measures; Mediating; Membrane Proteins; Molecular; Monitor; Mus; Organ; Organ Harvestings; Peptide/MHC Complex; Plasmids; Pneumonia; Population; Procedures; Process; Production; Protein Binding; Proteins; Radioactivity; Rate; Recombinants; Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Streptococcus pneumoniae; System; T-Cell Activation; T-Lymphocyte; Techniques; Testing; Toxic effect; Transgenic Organisms; Vaccinated; Vaccines; Work; aluminum sulfate; cost; cytokine; enzyme linked immunospot assay; in vivo; infectious disease model; pathogen; receptor; respiratory; response; vaccine delivery

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.

需要目前人类佐剂的替代方案。我们的研究表明，在没有 辅助，将抗原（Ag）靶向人类FCGAMMA受体I型（HFCGAMMARI）在Ag呈现的细胞上 在体内增强T细胞的体外和Ag特异性抗体（AB）和细胞因子的产生。然而， 没有人创建重组免疫原，可以检查HFCGAMMARI靶向 在传染病模型中的方法，或对所涉及机制的故意研究。链 肺炎球菌是一种呼吸道病原体，已鉴定出保护性AG（PSPA）。 PSPA是 细菌表面蛋白，当用作佐剂的免疫原时，会产生针对 小鼠肺炎链球菌感染。我们假设：1）可以保护肺炎链球菌感染 通过用抗Hfcgammari-PSPA融合蛋白免疫（在没有佐剂的情况下）增强（在没有佐剂）中增强。 2） 增强的机制还将涉及Ag定位到淋巴组织的改变 作为AG加工的改变。为了测试后者，我们将生成抗Hfcgammari-Hel融合蛋白。 HEL是一种通常用于进行Ag加工和表现的机械研究的蛋白质AG， 体内和体内。在Speific AIM 1中，我们将：a）抗抗Hfcgammari-PSPA和抗Hfcgammari- HEL Fusion蛋白已经生成。 b）检查融合蛋白增强T和B细胞的能力 体外和体内反应。 c）在体内确定hfcgammari ag靶向Ag的作用部分是通过 Ag的定位增强了淋巴组织。在特定目标2中，我们将：a）测试抗抗的能力 HFCGAMMARI-PSPA融合蛋白可预防肺炎链球菌的挑战。 b）研究影响 使用生化，免疫学和 超微结构技术。拟议的研究将为进一步的HFCGAMMARI靶向奠定基础 使用小鼠和人类中各种传染病药物的研究，消除了对传统的需求 佐剂，大大减少了接种个人所需的农业量（降低成本和 潜在的毒性），并提供疫苗输送系统，同时增强体液和 细胞免疫反应。因此，这些研究和该疫苗策略将适用于疫苗 针对成人，小儿和免疫功能低下的种群中各种各样的病原体。