Recombinant Subunit Protein for Flu Vaccine

流感疫苗用重组亚基蛋白

• 批准号: 6933266
• 负责人: Carolyn Louise Weeks-Levy
• 金额: $ 48.75万
• 依托单位: HAWAII BIOTECH, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-01 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6933266
• 关键词: Baculoviridae; Bombycidae; Drosophilidae; hemagglutinin; immune response; influenza vaccines; influenzavirus A; laboratory mouse; molecular cloning; protein purification; recombinant proteins; transfection; vaccine development; vaccine evaluation; virus protein

DESCRIPTION (provided by applicant): Current influenza vaccine production procedures (use of embryonated chicken eggs) inherently limit the amount of influenza vaccine that can be produced prior to each year's flu season. Most of the influenza vaccine sold is inactivated. Inactivation of the virus is accomplished through the use of agents such as formalin which is a compound that is known to cross-link protein and damage epitopes. There is a clear need for new technologies that can be used to respond quickly to influenza outbreaks and pandemics and produce sufficient doses of high quality vaccine for the U.S. population. There is also need for improved vaccine formulations with increased immunogenicity and efficacy. The overall goal of this research proposal is to evaluate recombinant subunit proteins of the avian influenza strain A/Hong Kong/156/97 (H5N1) as vaccine candidates. Recombinant subunits of the HA, NP and M1 proteins will be expressed in the Drosophila S2 expression system. The recombinantly expressed HA, NP and M1 proteins will be highly purified by column chromatography. The quality of the Drosophila and Bomftyx-expressed proteins is anticipated to be superior to conventionally produced influenza proteins due to the fact that the protein will not be altered by formalin or other agents used for inactivation which can cross-link proteins and destroy epitopes. The subunit proteins will be purified and tested in different combinations as vaccine immunogens in a mouse immunogenicity model. The proteins will also be expressed in the Bombyx mori system to provide Hawaii Biotech Inc an alternative expression system to the Drosophila system. Humoral and cellular immune responses will be evaluated in the mouse model. Dose ranging studies and different adjuvants will also be evaluated. The Drosophila and Bombyx mori expressed HA proteins will be compared to commercially available baculovirus expressed HA protein in the mouse immunogenicity model. Specific optimized vaccine formulations will then be evaluated in a mouse challenge model. In addition, the role of NP and M1 proteins in cellular immune responses will be explored.

描述（由申请人提供）：当前的流感疫苗生产程序（使用含胚鸡蛋）本质上限制了每年流感季节之前可以生产的流感疫苗的数量。大多数出售的流感疫苗都是灭活的。病毒的灭活是通过使用诸如福尔马林之类的试剂来实现的，福尔马林是一种已知能够交联蛋白质并破坏表位的化合物。显然需要新技术来快速应对流感爆发和大流行，并为美国民众生产足够剂量的高质量疫苗。还需要具有增加的免疫原性和功效的改进的疫苗制剂。该研究计划的总体目标是评估作为候选疫苗的禽流感病毒株 A/Hong Kong/156/97 (H5N1) 的重组亚基蛋白。 HA、NP 和 M1 蛋白的重组亚基将在果蝇 S2 表达系统中表达。重组表达的HA、NP和M1蛋白将通过柱层析得到高度纯化。果蝇和 Bomftyx 表达的蛋白质的质量预计优于传统生产的流感蛋白质，因为该蛋白质不会被福尔马林或用于灭活的其他试剂改变，这些试剂可以交联蛋白质并破坏表位。亚基蛋白将被纯化并以不同的组合作为小鼠免疫原性模型中的疫苗免疫原进行测试。这些蛋白质还将在家蚕系统中表达，为夏威夷生物技术公司提供果蝇系统的替代表达系统。将在小鼠模型中评估体液和细胞免疫反应。还将评估剂量范围研究和不同的佐剂。将在小鼠免疫原性模型中将果蝇和家蚕表达的 HA 蛋白与市售杆状病毒表达的 HA 蛋白进行比较。然后将在小鼠攻击模型中评估特定的优化疫苗配方。此外，还将探讨 NP 和 M1 蛋白在细胞免疫反应中的作用。