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：
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
项目类别：
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

项目摘要

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.

需要现有人类佐剂的替代品。我们的研究表明，在没有佐剂，将抗原 (Ag) 靶向 Ag 呈递细胞上的人 Fcgamma 受体 I 型 (hFcgammaRI) 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，没有人创造出重组免疫原，可以检查 hFcgammaRI 靶向传染病模型中的方法，或对所涉及机制的刻意研究。链球菌- 肺炎球菌是一种呼吸道病原体，已鉴定出具有保护性的 Ag (PspA)。 PspA 是一种细菌表面蛋白，当与佐剂一起用作免疫原时，可产生针对小鼠肺炎链球菌感染。我们假设：1）针对肺炎链球菌感染的保护作用可以是通过用抗 hFcgammaRI-PspA 融合蛋白进行免疫增强（在没有佐剂的情况下）。 2）增强的机制也将涉及 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 是一种蛋白质 Ag，通常用于进行 Ag 加工和呈现的机制研究，在体外和体内。在具体目标 1 中，我们将： A) 将抗 hFcgammaRI-PspA 和抗 hFcgammaRI-去人性化 HEL 融合蛋白已生成。 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 Ag 的 hFcgammaRI 靶向是否有效，部分通过增强Ag在淋巴组织中的定位。在具体目标 2 中，我们将： A) 测试抗- hFcgammaRI-PspA 融合蛋白可防止肺炎链球菌的攻击。 B) 研究影响 hFcgammaRI 利用生化、免疫学和超微结构技术。拟议的研究将为进一步的 hFcgammaRI 靶向奠定基础在小鼠和人类身上使用多种传染病病原体的研究消除了传统治疗的需要佐剂，大幅减少个人疫苗接种所需的 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疫苗针对成人、儿童和免疫功能低下人群的多种病原体。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Mucosal immunization with an unadjuvanted vaccine that targets Streptococcus pneumoniae PspA to human Fcγ receptor type I protects against pneumococcal infection through complement- and lactoferrin-mediated bactericidal activity.

使用针对人 Fcγ 受体 I 型的肺炎链球菌 PspA 的无佐剂疫苗进行粘膜免疫，可通过补体和乳铁蛋白介导的杀菌活性来防止肺炎球菌感染。

DOI：
发表时间：
2012-03
期刊：
影响因子：
3.1
作者：
Bitsaktsis, Constantine;Iglesias, Bibiana V;Li, Ying;Colino, Jesus;Snapper, Clifford M;Hollingshead, Susan K;Pham, Giang;Gosselin, Diane R;Gosselin, Edmund J
通讯作者：
Gosselin, Edmund J