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性脑炎。它在整个亚洲都很普遍，并正在超越其传统边界。没有专门的治疗方法。目前，在不同国家使用的乙脑疫苗有3种，但国际上只有一种，即小鼠脑源性灭活疫苗。虽然这种疫苗有效地降低了乙脑的发病率，但它相对昂贵，并且与严重的过敏和神经反应有关。其他2种乙脑疫苗因监管问题仅在中国使用。乙脑疫苗的开发是世界卫生组织（WHO）的一项高度优先事项。迄今为止，还没有研究采用复制缺陷型JEV病毒粒子作为免疫原来刺激有效的免疫。我们最近开发了一种高效的DNE 2/AC复制子包装细胞系。假感染性病毒样颗粒（PVLP）在BHK-21细胞上的传代表明，即使在包装细胞上传代6次后，也完全不存在任何感染性病毒。因此，为了开发更安全、更有效、更廉价的乙脑疫苗，我们构建了乙脑病毒（JEV）SA 14 -14-2株的全长感染性cDNA克隆。基于这些非常有希望的结果，本I期SBIR建议的具体目标是：1）构建JEV SA 14 -14-2株的JEV/AC复制子; 2）开发稳定的包装细胞系，其在培养中提供JE结构蛋白C; 3）从感染的包装细胞系收获JEV PVLP，并分析包装细胞系和PVLP在传代过程中的稳定性; 4）对拟定疫苗在小鼠中的免疫原性进行初步分析。该项目的成功完成将使SBIR II期研究得以进行，包括开发工业规模生产PVLP的具有成本效益的GMP工艺，体内免疫原决定因素的滴定，对PVLP的体液和细胞反应的评估，以及对乙脑感染免疫力的临床前动物研究。 假感染性病毒样颗粒作为疫苗候选物是安全性和有效性的内在最佳组合。低成本生产对于实际应用至关重要。在本提案中，我们将开发一种可用于大规模生产的高效包装系统。