Development of Pseudoinfectious VLP as Novel JEV Vaccine

新型乙脑疫苗假传染性VLP的研制

• 批准号: 7280433
• 负责人: XIAOWU PANG
• 金额: $ 28.45万
• 依托单位: TENGEN BIOMEDICAL CO.
• 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 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7280433
• 关键词: Acute; Adult; Affect; Agriculture; Allergic; Allergic Reaction; Animals; Antigens; Antiviral Agents; Area; Asia; Asians; Attenuated; Australia; Brain; Cell Line; Cells; Central Nervous System Viral Diseases; Child; China; Complementary DNA; Country; Culicidae; DNA; Dengue; Dengue Virus; Depth; Development; Disabled Persons; Disease; Domestic Animals; Dose; Essential Genes; Facility Construction Funding Category; Far East; Flavivirus; Formalin; Foundations; Genes; Genome; Geographic Locations; Goals; Guanosine Monophosphate; Harvest; Human; Immunity; In Vitro; Inactivated Vaccines; Incidence; India; Indium; Infection; Infectious Agent; Interferon-alpha; International; Japan; Japanese Encephalitis; Japanese Encephalitis Vaccines; Japanese encephalitis virus; Knowledge; Korea; Length; Lethal Dose 50; Life; Link; Location; Measurable; Measures; Mus; Neurologic; Outcome; Pharmaceutical Preparations; Phase; Process; Production; Protein C; Proteins; Reaction; Replicon; Reporting; Research; Risk; Safety; Serotyping; Small Business Funding Mechanisms; Small Business Innovation Research Grant; Staging; Standards of Weights and Measures; Structural Protein; Subunit Vaccines; Survivors; System; Titrations; Training; Vaccination; Vaccines; Viral Encephalitis; Virion; Virus; Virus Diseases; Virus-like particle; World Health Organization; Yellow Fever; base; cost; double-blind placebo controlled trial; genetic manipulation; immunogenicity; in vivo; novel; pathogen; pre-clinical; programs; recombinant virus; response; success; vaccine development

DESCRIPTION (provided by applicant): Japanese encephalitis (JE) is the most important viral encephalitis in the world. It is widespread throughout Asia and is spreading beyond its traditional boundaries. There is no specific treatment for JE. Currently, 3 kinds of JE vaccine are in use in different countries, but only one is available internationally, a mouse-brain-derived inactivated vaccine. Although, this vaccine has been effective in reducing the incidence of JE, it is relatively expensive and has been linked to severe allergic and neurological reactions. Other 2 JE vaccines are only used in China, due to regulatory concern. Vaccine development for JE is a high priority on the list of World Health Organization (WHO). To date, no study has employed a replication-incompetent JEV virion as immunogens to stimulate effective immunity. We have recently developed a highly efficient packaging cell line for DNE2/AC replicon. Passaging of the pseudoinfectious virus-like particles (PVLP) on BHK-21 cell illustrated the complete absence of any infectious virus, even after 6 passages on packaging cells. Thus, toward the overall goal of developing a safer, more effective, and less costly JE vaccine, we have constructed a full-length infectious cDNA clone for JE virus (JEV) SA14-14-2 strain. Based upon these highly promising results, the Specific Aims of this Phase I SBIR proposal are: 1) construction of JEV/AC replicon for JEV SA14-14-2 strain; 2) development of stable packaging cell lines providing JE structural protein C in trains; 3) harvest of JEV PVLPs from infected packaging cell line, and analyzing the stability of the packaging cell lines and PVLP during passages; 4) preliminary analysis of the immunogenicity of the proposed vaccine in mice. Successful completion of this program will enable SBIR Phase II research including development of a cost-effective GMP process for PVLP production at industrial scale, titration of the determinants as immunogens in vivo, assessment of both the humoral and cellular responses to the PVLP, and a pre-clinical animal study of immunity to JE infection. Pseudoinfectious virus-like particle as vaccine candidate is inherent optimal combination of safety and efficacy. Low cost production is critical for practical use. In this proposal, we are going to develop a high efficient packaging system that can be used for large scale production.

描述（由申请人提供）：日本脑炎（JE）是世界上最重要的病毒脑炎。它在整个亚洲都是广泛的，并且正在超越其传统界限。没有针对JE的特定治疗方法。目前，在不同国家使用了3种JE疫苗，但在国际上只有一种可用的小鼠脑源性疫苗。尽管该疫苗可有效降低JE的发生率，但它相对昂贵，并且与严重的过敏和神经反应有关。由于监管问题，其他2种JE疫苗仅在中国使用。 JE的疫苗开发是世界卫生组织（WHO）列表中的高度优先事项。迄今为止，尚无研究用不含复制的JEV病毒剂作为免疫原子来刺激有效的免疫力。我们最近为DNE2/AC复制子开发了高效的包装细胞系。 BHK-21细胞上假病毒样颗粒（PVLP）的传播表明，即使在包装细胞上进行了6次通道后，也完全没有任何感染性病毒。因此，为了开发一种更安全，更有效且成本较低的JE疫苗的总体目标，我们为JE病毒（JE）SA14-14-2菌株构建了全长的感染性cDNA克隆。基于这些高度有希望的结果，该阶段I SBIR提案的具体目的是：1）JEV SA14-14-2菌株的JEV/AC复制子的构建； 2）开发稳定的包装细胞系，在火车中提供JE结构蛋白C； 3）从受感染的包装细胞系中收获JEV PVLP，并分析通道过程中包装细胞系和PVLP的稳定性； 4）对小鼠提出的疫苗的免疫原性的初步分析。该计划的成功完成将使SBIR II期研究能够开发以工业规模的PVLP生产成本有效的GMP流程，将决定因素作为体内免疫原子的滴定，评估对PVLP的体液和细胞反应以及对PVLP的体液和细胞反应，以及对JE感染的免疫动物研究。 伪感染病毒样颗粒作为疫苗候选者是安全性和功效的最佳组合。低成本生产对于实际使用至关重要。在此提案中，我们将开发一个可用于大规模生产的高效包装系统。