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.

目前人类佐剂的替代品是必要的。我们的研究表明，在没有 人FcGammaI型受体(HFcGammaRI)佐剂靶向抗原在抗原提呈细胞上的表达 在体外增强T细胞活化，在体内增强抗原特异性抗体和细胞因子的产生。然而， 还没有人创造出能够检测hFcGammaRI靶向的重组免疫原 传染病模型中的方法，或对所涉及的机制的刻意研究(S)。斯特雷普托- 肺炎球菌是一种呼吸道病原体，其保护性抗原(PSPA)已被鉴定。PSPA是 一种细菌表面蛋白，当作为免疫原与佐剂一起使用时，可产生对 小鼠肺炎链球菌感染。我们假设：1)对肺炎链球菌感染的保护措施 通过用抗hFcGammaRI-PSPA融合蛋白免疫增强(在没有佐剂的情况下)。2) 增强的机制(S)也将涉及到淋巴组织中Ag定位的改变 作为银加工过程中的蚀变。为了测试后者，我们将产生一个抗hFcGammaRI-HEL融合蛋白。 HELs是一种蛋白质Ag，通常用于进行Ag加工和呈递的机制研究，在 体外和体内。在特殊目标1中，我们将：a)使抗hFcGammaRI-PSPA和抗hFcGammaRI非人性化- 已经产生了HELL融合蛋白。B)检查融合蛋白增强T和B细胞的能力 体外和体内的反应。C)在体内确定hFcGammaRI靶向Ag是否部分通过 Ag在淋巴组织中的定位增强。在具体目标2中，我们将：a)测试抗病毒能力 HFcGammaRI-PSPA融合蛋白保护肺炎链球菌的攻击。B)研究其影响 HFcGammaRI靶向抗原加工和呈递利用生化、免疫学和 超微结构技术。建议的研究将为进一步的hFcGammaRI靶向奠定基础 在老鼠和人类身上使用多种传染病病原体的研究，消除了传统的 佐剂，大大减少个人接种疫苗所需的银量(降低成本和 潜在毒性)，并提供一种疫苗递送系统，同时增强体液和 细胞免疫反应。因此，这些研究和疫苗策略将适用于疫苗。 在成人、儿童和免疫受损人群中对抗各种各样的病原体。