Vaccine Development against Infectious Diseases

传染病疫苗的开发

• 批准号: 9242651
• 负责人: Miguel Otero
• 金额: $ 11.25万
• 依托单位: UNIVERSITY OF PUERTO RICO MED SCIENCES
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-03-11 至 2020-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9242651
• 关键词: Address; Adjuvant; Antibodies; Antigen Presentation; Antigen Targeting; Antigen-Presenting Cells; Antigens; Antiviral Agents; Attenuated Vaccines; Binding Proteins; Biological Response Modifiers; Bioterrorism; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CD8B1 gene; Cell Proliferation; Cells; Chimera organism; Clinic; Code; Codon Nucleotides; Collaborations; Communicable Diseases; DNA; DNA Vaccines; Data; Dendritic Cells; Disease; Dose; Ebola-like Viruses; Engineering; Enterovirus 68; Enzyme-Linked Immunosorbent Assay; Frequencies; Generations; Genes; Glycoproteins; Goals; HIV; HIV Envelope Protein gp120; HIV-1; IgG1; Imiquimod; Immune; Immune response; Immune system; Immunity; Immunization; Immunize; Immunoglobulin G; Immunologic Adjuvants; Immunomodulators; Immunotherapy; In Vitro; Individual; Infection; Influenza; Interferon Type II; Lymphocyte; Mediating; Memory; Modeling; Morbidity - disease rate; Mus; Piedra; Plasmids; Polysaccharides; Population; Production; Property; Proteins; Protocols documentation; Public Health; Quality of life; Resources; Route; Serum; Smallpox; Spleen; Splenocyte; Stimulus; Surface Antigens; Testing; Therapeutic Intervention; Translating; Vaccination; Vaccine Design; Vaccines; Vaccinia virus; Viral; Viral Vector; Viremia; Virus; Virus Activation; Virus Replication; base; cell mediated immune response; chikungunya; cytokine; design; effective therapy; enzyme linked immunospot assay; immunogenic; immunoregulation; in vivo; interest; long term memory; lymphocyte proliferation; mortality; nanoparticle; neutralizing antibody; new therapeutic target; novel; novel strategies; pathogen; plasmid DNA; public health relevance; sugar; vaccine development; vector

 DESCRIPTION (provided by applicant): The concomitant danger and global spread of viruses like Ebola, Chikungunya, Enterovirus D68, and Influenza, makes a priority to develop effective vaccines against emerging diseases. In this regard, live vaccines have been effective resources against infections. However, they are deleterious for immune compromised individuals. Thus, there is a critical need for the production of safe virus-free vaccines. For that reason, our goal s to manipulate the mechanisms that regulate the immune responses, in order to generate safe vaccination approaches that could be broadly used by most of the population. In this regard, our preliminary data in collaboration with Dr. José Rivera (UPR-Río Piedras Campus), shows that a novel Supramolecular Nanoparticle (NP) has strong immunomodulator properties when tested in vivo using a DNA vaccination approach. Also, we characterized the immunogenic potential of a Coriolus versicolor polysaccharide (PSP) extract. Our in vitro data shows PSP to stimulate lymphocyte proliferation and to induce a TH1 cytokine profile. This strong preliminary data is very encouraging, and drove our interest to study the immunomodulatory effects of NP and PSP in a DNA vaccination approach. To the best of our knowledge, the immunomodulator activity of any of these novel agents has never been tested in a DNA vaccination regime. We engineered our model antigen to target Dendritic Cells (DCs), as these are potent antigen-presenting cells (APCs). Specifically, HIV-1 Outer Domain-1 (OD1) is a highly-mannosylated motif that routs HIV to Antigen Presenting Cells (APCs). For that reason we tested a plasmid DNA vaccine consisting of a codon-optimized chimera coding for OD1 fused by a linker sequence to the Vaccinia Virus (VACV) A27L antigenic protein to enhance antigen targeting to APCs. Our preliminary data shows that mice immunized with our plasmid (pOD1A27Lopt) produce higher A27-specific cell- and humoral-mediated immune responses, compared to controls. Based on this information, we hypothesize that immune responses produced by pOD1A27Lopt formulated in NP and PSP, will induce long-term immune protection. We will address our hypothesis with the following specific aims: Specific Aim 1. Dose-dependent immunomodulatory effect of our vaccine formulation in cell-mediated immune responses, lymphoproliferation, and cytokine profile. We will compare the effect of NP and PSP in the antigen-specific frequency of CD8+ and CD4+ T-cells releasing IFN-γ in spleen cells by ELISPOT analysis, and CD4+ T-cell proliferation. Also, we will study the cytokine profile in serum of immunized mice. Specific Aim 2. Dose-dependent immunomodulatory effect of our vaccine formulation in humoral- mediated immune responses. We will compare the effect of NP and PSP in the antigen-specific production of total IgG, IgG1 (Th2-type), and IgG2a (Th1-type) by ELISA, and the generation of neutralizing antibodies. Specific Aim 3. Immunomodulatory effect of our vaccine formulation in viral replication, memory, activation, and proliferation. We will compare the effect of NP and PSP in the enhancement of antigen-specific protective immunity with a viral challenge after immunization. Also, we will determine the effect of this vaccination protocol on viral replication and induction of long-term memory, activation and proliferation. We expect our vaccine formulation to offer novel approaches for rational-vaccine design against emerging diseases and bioterror agents, advancing the field of immunotherapy. This project will provide new targets for therapeutic interventions to induce safe immune protection.

 描述（由申请人提供）：埃博拉病毒、基孔肯雅病毒、肠道病毒 D68 和流感等病毒的伴随危险和全球传播，使得开发针对新出现疾病的有效疫苗成为首要任务。在这方面，活疫苗是对抗感染的有效资源。然而，它们对免疫受损的个体是有害的，因此，迫切需要生产安全的无病毒疫苗，因此，我们的目标是操纵调节免疫反应的机制。为了制定可供大多数人广泛使用的安全疫苗接种方法，我们与 José Rivera 博士（UPR-Río Piedras 校区）合作的初步数据表明，一种新型超分子纳米颗粒 (NP) 具有以下特点：使用 DNA 疫苗接种方法进行体内测试时，我们还鉴定了云芝多糖 (PSP) 提取物的免疫原性潜力。我们的体外数据显示，PSP 可以刺激淋巴细胞增殖。并诱导 TH1 细胞因子谱，这一强有力的初步数据非常令人鼓舞，并激发了我们研究 NP 和 PSP 在 DNA 疫苗接种方法中的免疫调节作用的兴趣。我们的模型抗原从未在 DNA 疫苗接种方案中进行过测试，因为它们是有效的抗原呈递细胞 (APC)。因此，我们测试了一种质粒 DNA 疫苗，该疫苗由密码子优化的编码 OD1 的嵌合体组成，该嵌合体通过连接序列与痘苗病毒 (VACV) A27L 抗原蛋白融合。我们的初步数据表明，用我们的质粒 (pOD1A27Lopt) 免疫的小鼠产生更高的 A27 特异性细胞-与对照组相比，我们发现 pOD1A27Lopt 产生的免疫反应将诱导长期免疫保护，具体目标如下：目标 1. 我们的疫苗制剂在细胞介导的免疫反应、淋巴细胞增殖和细胞因子谱中的剂量依赖性免疫调节作用我们将比较 NP 和 PSP 对 CD8+ 和 CD8+ 的抗原特异性频率的影响。通过 ELISPOT 分析，CD4+ T 细胞在脾细胞中释放 IFN-γ，以及 CD4+ T 细胞增殖。此外，我们还将研究免疫小鼠血清中的细胞因子谱。 我们的疫苗制剂在体液介导的免疫反应中的剂量依赖性免疫调节作用我们将比较 NP 和 PSP 在总 IgG、IgG1（Th2 型）和 IgG2a（Th1 型）的抗原特异性产生中的效果。 ELISA，以及中和抗体的产生。 具体目标 3. 我们的疫苗制剂对病毒复制、记忆、激活和增殖的免疫调节作用 我们将比较 NP 和 PSP 在病毒复制、记忆、激活和增殖中的作用。免疫后通过病毒攻击增强抗原特异性保护性免疫力此外，我们将确定该疫苗接种方案对病毒复制和诱导长期记忆、激活和增殖的影响。针对新出现的疾病和生物恐怖分子的合理疫苗设计，推进了免疫治疗领域的发展，该项目将为治疗干预提供新的目标，以诱导安全的免疫保护。