Protective Activity of a Multi-Functional Immunogen

多功能免疫原的保护活性

基本信息

批准号：
7195315
负责人：
Edmund J Gosselin
金额：
$ 19.75万
依托单位：
ALBANY MEDICAL COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-02-15 至 2009-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7195315
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

项目摘要

DESCRIPTION (provided by applicant): 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.

描述（由申请人提供）：需要现有人类佐剂的替代品。我们的研究表明，在没有佐剂的情况下，将抗原 (Ag) 靶向 Ag 呈递细胞上的人 Fcgamma 受体 I 型 (hFcgammaRI) 可增强体外 T 细胞活化以及体内 Ag 特异性抗体 (Ab) 和细胞因子的产生。然而，还没有人创造出重组免疫原，从而可以在传染病模型中检查 hFcgammaRI 靶向方法，或对所涉及的机制进行深思熟虑的研究。肺炎链球菌是一种呼吸道病原体，已鉴定出具有保护作用的 Ag (PspA)。 PspA 是一种细菌表面蛋白，当其与佐剂一起用作免疫原时，可以在小鼠体内产生针对肺炎链球菌感染的保护作用。我们假设：1) 通过使用抗 hFcgammaRI-PspA 融合蛋白进行免疫可以增强对肺炎链球菌感染的保护（在没有佐剂的情况下）。 2) 增强机制将涉及Ag定位到淋巴组织的改变，以及Ag处理的改变。为了测试后者，我们将生成抗 hFcgammaRI-HEL 融合蛋白。 HEL 是一种蛋白质 Ag，通常用于在体外和体内进行 Ag 加工和呈递的机制研究。在具体目标 1 中，我们将： A) 将已生成的抗 hFcgammaRI-PspA 和抗 hFcgammaRI-HEL 融合蛋白去人性化。 B) 检查融合蛋白在体外和体内增强 T 细胞和 B 细胞反应的能力。 C) 在体内确定 Ag 的 hFcgammaRI 靶向是否部分通过增强 Ag 到淋巴组织的定位而起作用。在具体目标 2 中，我们将： A) 测试抗 hFcgammaRI-PspA 融合蛋白抵御肺炎链球菌攻击的能力。 B) 利用生化、免疫学和超微结构技术研究 hFcgammaRI 靶向对 Ag 加工和呈递的影响。拟议的研究将为在小鼠和人类中使用多种传染病因子进行进一步的 hFcgammaRI 靶向研究奠定基础，消除对传统佐剂的需求，大幅减少个体疫苗接种所需的 Ag 量（降低成本和潜在毒性）），并提供疫苗输送系统，同时增强体液和细胞免疫反应。因此，这些研究和这种疫苗策略将适用于针对成人、儿童和免疫功能低下人群的多种病原体的疫苗。