Vaccine Development against Infectious Diseases

传染病疫苗的开发

• 批准号: 8998543
• 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/8998543
• 关键词: Address; Adjuvant; Antibodies; Antigen Presentation; Antigen Targeting; Antigen-Presenting Cells; Antigens; Antiviral Agents; Attenuated Vaccines; Binding Proteins; Biological Response Modifiers; CD4 Positive T Lymphocytes; CD8B1 gene; 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; Immunoglobulin G; Immunologic Adjuvants; Immunomodulators; Immunotherapy; In Vitro; Individual; Infection; Influenza; Interferon Type II; Lead; Lymphocyte; Mediating; Memory; Modeling; Morbidity - disease rate; Mus; Piedra; Plasmids; Polysaccharides; Population; Production; Property; Proteins; Protocols documentation; Public Health; Quality of life; Resources; Serum; Smallpox; Spleen; Splenocyte; Stimulus; Surface Antigens; T-Cell Proliferation; T-Lymphocyte; 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; in vivo; interest; long term memory; lymphocyte proliferation; mortality; nanoparticle; neutralizing antibody; 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（由适用提供）：埃博拉病毒，基孔肯尼亚，肠病毒D68和流感等病毒的危险和全球蔓延，优先考虑开发针对新兴疾病的有效疫苗。在这方面，现场疫苗是针对感染的有效资源。但是，他们对免疫人员组成的个体是有害的。这是对生产无病毒疫苗的迫切需要。因此，我们的目标是操纵调节免疫调查的机制，以生成大多数人群可以广泛使用的安全疫苗方法。在这方面，我们与JoséRivera博士（UPR-RíoPiedras校园）合作的初步数据表明，新型的超分子纳米颗粒（NP）使用DNA疫苗方法进行体内测试时具有强大的免疫调节剂。此外，我们表征了科里奥鲁斯剂量多糖（PSP）提取物的免疫原性潜力。我们的体外数据显示PSP刺激淋巴细胞增殖并诱导Th1细胞因子谱。这种强大的初步数据非常令人鼓舞，并引起了我们研究NP和PSP在DNA疫苗方法中的免疫调节作用的兴趣。据我们所知，这些新型药物中任何一种的免疫调节剂活性从未在DNA疫苗状态下进行测试。我们将模型抗原设计为靶向树突状细胞（DC），因为它们是潜在的抗原呈递细胞（APC）。具体而言，HIV-1外域-1（OD1）是一种高度曼诺糖基化的基序，将HIV与抗原呈现细胞（APC）相关。因此，我们测试了由密码子优化的嵌合体组成的质粒DNA疫苗，该嵌合体为vacinia病毒（VACV）A27L抗原蛋白融合的OD1编码，以增强抗原靶向APC。我们的初步数据表明，与对照组相比，我们的质粒（POD1A27LOPT）免疫了小鼠（POD1A27LOPT）可产生较高的A27特异性细胞和体液介导的免疫血液。基于这些信息，我们假设由NP和PSP制定的POD1A27LOPT产生的免疫复杂将诱导长期免疫力。我们将以以下特定目的来解决我们的假设：特定目标1。疫苗配方在细胞介导的免疫调查中，淋巴增生和细胞因子谱的剂量依赖性免疫调节作用。我们将通过ELISPOT分析以及CD4+ T细胞增殖来比较NP和PSP在脾细胞中释放IFN-γ的CD8+和CD4+ T细胞的抗原特异性频率。此外，我们将研究免疫抑制小鼠血清中的细胞因子谱。具体目标2。 我们的疫苗配方中剂量依赖性免疫调节作用在体液介导的免疫调查中。我们将比较NP和PSP对ELISA的总IgG，IgG1（Th2-Type）和IgG2A（Th1型）的抗原特异性产生的影响，以及中和抗体的产生。特定目的3。疫苗配方在病毒复制，记忆，激活和增殖中的免疫调节作用。我们将比较NP和PSP在免疫后使用病毒挑战的NP和PSP对增强抗原特异性免疫学的影响。此外，我们将确定该疫苗方案对长期记忆，激活和增殖的病毒复制和诱导的影响。我们希望我们的疫苗配方为针对新兴疾病和生物疾病剂的理性疫苗设计提供新颖的方法，从而推进免疫疗法领域。该项目将为治疗干预措施提供新的目标，以诱导安全免疫学保护。