Dev. of Optimized/Adjuvanted Smallpox DNA Vaccine (Gene Gun)

开发。

• 批准号: 7281724
• 负责人: TZYY-CHOOU WU
• 金额: $ 30.33万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7281724
• 关键词: Adjuvant; Adverse event; Animal Model; Antibody Formation; Antigen Presentation; Antigen Presentation Pathway; Antigen-Presenting Cells; Antigens; Antiviral Agents; Attenuated; Biological; Biological Assay; Body Weight decreased; Boxing; Cellular Immunity; Chemical Vaccines; Clinical; Clinical Trials; Codon Nucleotides; DNA; DNA Vaccines; DNA delivery; Data; Development; Devices; Drug Formulations; Enterotoxins; Enzyme-Linked Immunosorbent Assay; Enzymes; Flow Cytometry; Genes; Genetic; Granulocyte-Macrophage Colony-Stimulating Factor; HLA-A2 Antigen; Hand functions; Heating; Immune; Immune response; Immunity; Infection; Intradermal Tests; Intravenous; Lead; Letters; Licensing; Life; Link; Macaca; Macaca mulatta; Measures; Membrane Proteins; Methods; Military Personnel; Modified Vaccinia Virus Ankara; Monkeypox; Monkeypox virus; Monoclonal Antibodies; Mus; ND-10; Numbers; Panic; Persons; Plasmids; Play; Population; Poxviridae; Poxviridae Infections; Primate Diseases; Proteins; Radioimmunoprecipitation Assay; Range; Relative (related person); Research Personnel; Risk; Role; Route; Smallpox; Smallpox Vaccine; Spottings; Staining method; Stains; T-Lymphocyte; Testing; Transgenic Mice; United States; Vaccinated; Vaccination; Vaccines; Vaccinia virus; Virus; Virus Diseases; Western Blotting; Work; base; calreticulin; cell mediated immune response; cytokine; enterotoxigenic Escherichia coli; gene gun; immunogenicity; improved; in vivo; interest; neutralizing antibody; nonhuman primate; novel; novel vaccines; programs; response; vaccination strategy; vector control

DESCRIPTION (provided by applicant): The reemergence of smallpox (or derivative) in an unprotected civilian or military population would likely cause mass casualties, panic, and functional breakdown. Thus, the availability of a safe effective poxvirus vaccine may significantly alleviate some of these concerns. Among the poxvirus-encoded proteins, four membrane proteins (A33R, B5R, L1R and A27L) (called 4-pox vaccine) have been shown to generate protective neutralizing antibody responses and host range protein 2 (HRP2, also called 018L) has been shown to generate protective T cell-mediated immune response against poxvirus challenge in HLA-A2 transgenic mice. DNA vaccines targeting these key smallpox antigens represent a potentially plausible approach for the control of smallpox infections. A limitation of this DNA vaccination strategy is the naturally low immunogenicity of DNA encoding antigen, resulting in limited vaccine potency. Intradermal administration of DNA vaccines via gene gun represents an efficient method for delivery of DNA vaccines into professional APCs in vivo. The focus of the present application is to investigate different methods of enhancing potency based upon strategies that target the APCs. By selecting five methods of immune enhancement that rely on different mechanisms of improving antigen presentation, it is hoped that novel additive or synergistic effects will be elicited to generate effective immunity. These methods include: 1) codon optimization of the 4-pox antigens to enhance antigen expression; 2) linkage of poxvirus antigen to calreticulin (CRT) to enhance antigen processing and presentation; 3) co-administration with DNA encoding heat labile enterotoxin (LT) from enterotoxigenic E. coli to enhance immunogenicity; 4) co-administration with a plasmid expressing GM-CSF to attract APCs and adjuvant DNA vaccines; and 5) addition of a DNA vaccine encoding a documented target protein of cell-mediated immunity (i.e. poxvirus HRP2 protein). Thus, in the current application, we will test if intradermal delivery of a codon-optimized 4-pox DNA vaccines and/or HRP2 DNA adjuvanted by inclusion of GM-CSF or LT DNA and/or linkage to CRT via gene gun would lead to enhanced antigen-specific humoral and T cell-mediated immune responses and antiviral effects against live poxvirus infections in vivo. The successful implementation of the proposed studies will result in an improved poxvirus DNA vaccine potentially suitable for clinical development.

描述（由申请人提供）：天花（或衍生物）在未受保护的平民或军事人群中的重新出现可能会导致大规模伤亡，恐慌和功能崩溃。因此，安全有效的痘病毒疫苗的可用性可能会显着减轻这些问题中的一些。在痘病毒编码的蛋白中，四种膜蛋白（A33 R、B5 R、L1 R和A27 L）（称为4痘疫苗）已被证明产生保护性中和抗体应答，并且宿主范围蛋白2（HRP 2，也称为018 L）已被证明在HLA-A2转基因小鼠中产生针对痘病毒攻击的保护性T细胞介导的免疫应答。针对这些关键天花抗原的DNA疫苗代表了控制天花感染的潜在可行方法。这种DNA疫苗接种策略的限制是编码抗原的DNA的天然低免疫原性，导致有限的疫苗效力。基因枪皮内注射DNA疫苗是将DNA疫苗导入体内专职APC的有效方法。本申请的重点是研究基于靶向APC的策略增强效力的不同方法。通过选择依赖于改善抗原呈递的不同机制的五种免疫增强方法，希望引起新的累加或协同效应以产生有效的免疫。这些方法包括：1）对4-痘抗原进行密码子优化，以增强抗原表达; 2）将痘病毒抗原与钙网蛋白（CRT）连接，以增强抗原加工和提呈; 3）与编码来自产肠毒素大肠杆菌的热不稳定肠毒素（LT）的DNA共同给药。coli增强免疫原性; 4）与表达GM-CSF的质粒共同给药以吸引APC和佐剂DNA疫苗;和5）添加编码已记录的细胞介导免疫靶蛋白（即痘病毒HRP 2蛋白）的DNA疫苗。因此，在本申请中，我们将测试通过包含GM-CSF或LT DNA和/或经由基因枪与CRT连接而佐剂化的密码子优化的4-痘DNA疫苗和/或HRP 2 DNA的皮内递送是否会导致增强的抗原特异性体液和T细胞介导的免疫应答以及针对活痘病毒感染的体内抗病毒作用。拟议研究的成功实施将导致改进的痘病毒DNA疫苗可能适用于临床开发。