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）：需要当前人类佐剂的替代方案。我们的研究表明，在没有佐剂的情况下，将抗原（Ag）靶向Ag呈现细胞的I型I型I型受体I（HFCGAMMARI）在体内增强了T细胞的体外和Ag特异性抗体（AB）和细胞因子的产生。然而，没有人会创建重组免疫原，可以在传染病模型中检查HFCGAMMARI靶向方法，或者对所涉及的机制进行故意研究。肺炎链球菌是一种呼吸道病原体，已鉴定出保护性AG（PSPA）。 PSPA是一种细菌表面蛋白，当该蛋白用作辅助的免疫原时，会在小鼠中产生针对肺炎链球菌感染的保护。我们假设：1）通过用抗HFCGAMMARI-PSPA融合蛋白免疫来增强对肺炎链球菌感染的保护。 2）增强的机制将涉及AG定位到淋巴组织的改变以及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是一种通常用于在体外和体内进行Ag加工和表现的机械研究的蛋白质AG。在Speific Aim 1中，我们将：a）已生成的抗Hfcgammari-PSPA和抗Hfcgammari-fusion蛋白。 b）检查融合蛋白在体外和体内增强T和B细胞反应的能力。 c）在体内确定hfcgammari ag的靶向部分，部分是通过增强Ag定位到淋巴组织的定位。在特定目标2中，我们将：a）测试抗HFCGAMMARI-PSPA融合蛋白防止肺炎链球菌挑战的能力。 b）研究靶向HFCGAMMARI对使用生化，免疫和超微结构技术的AG加工和表现的影响。拟议的研究将为使用小鼠和人类中各种传染病药物进行进一步的HFCGAMMARI靶向研究奠定基础，从而消除了对传统佐剂的需求，大大减少了疫苗接种所需的AG量（降低成本和潜在毒性），并提供疫苗输送系统，同时增强了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成人，小儿和免疫功能低下的种群中各种病原体的疫苗。