Developing Avirulent Rabies Virus Vaccines

描述：(申请人提供)狂犬病仍然是一种公共卫生 每年造成7万多人死亡的威胁。人类会被感染 狂犬病病毒主要通过家畜和野生动物的叮咬传播 动物。控制家畜和野生动物中的狂犬病病毒感染， 因此，不仅降低了这些动物的死亡率，而且还减少了 人类暴露的风险。为以下人群接种暴露前疫苗 持续处于危险中进一步预防人类狂犬病，暴露后也是如此 为被狂犬病或疑似狂犬病动物叮咬的人进行免疫接种。 目前，国内主要使用狂犬病灭活疫苗进行免疫 动物，尤其是宠物。纯化和灭活的狂犬病病毒疫苗有 用于人体在曝光前或曝光后的设置。重组牛痘疫苗 表达狂犬病病毒糖蛋白(VRG)的病毒已用于控制 野生动物中的狂犬病。虽然这些疫苗是有效的，但每年接种疫苗 需要对宠物保持足够的免疫力。对人类来说，多次服用 灭活的组织培养疫苗需要刺激最佳 免疫反应。此外，目前的组织培养疫苗价格昂贵； 因此，大多数需要接种疫苗的人(在发展中国家)不能 买得起它们。因此，有必要开发出更有效和负担得起的 狂犬病病毒疫苗。我们建议开发无毒的活狂犬病病毒 通过构建在体内传播能力降低的突变病毒来接种疫苗 神经系统(通过糖蛋白G的突变)和降低的 病毒复制(通过狂犬病内基因的突变和/或重排 病毒基因组)。这将通过使用最先进的反向 遗传学技术。我们的建议也是基于我们最近的调查结果。 因为其他人显示以下内容。1)磷酸化丝氨酸的突变 389%的N到丙氨酸使病毒复制速度降低了超过 五倍，病毒产量超过一万倍。2)G基因突变 At残基333显著降低狂犬病的毒力和致病性 病毒。3)水疱性口炎病毒内基因重排 基因组导致免疫应答的减弱和增强。这个 构建突变或重排狂犬病病毒的基本原理是 改变后的病毒很可能会比目前可用的更弱 减毒狂犬病病毒(仍能在新生动物中引发狂犬病)。如果 进一步减毒的狂犬病病毒不再在动物身上引起任何疾病 年龄和任何接种途径，但仍保持免疫力，它们可能是 开发为人和动物的改良活狂犬病疫苗。

DESCRIPTION: (Provided by Applicant) Rabies still presents a public health threat causing more than 70,000 human deaths each year. Humans get infected with the rabies virus mostly through bites from rabid domestic and wildlife animals. Controlling rabies virus infection in domestic and wildlife animals, therefore, not only reduces the mortality in these animals but also reduces the risks of human exposure. Pre-exposure vaccinations for people who are constantly at risk further prevent human rabies, as do post-exposure immunizations for people who are bitten by rabid or suspected rabid animals. Currently, inactivated rabies virus vaccines are used to immunize domestic animals, particularly pets. Purified and inactivated rabies virus vaccines are used for humans in the pre- or post-exposure settings. A recombinant vaccinia virus expressing rabies virus glycoprotein (VRG) has been used to control rabies in wildlife. Although these vaccines are effective, annual vaccinations are required to maintain adequate immunity in pets. For humans, multiple doses of the inactivated tissue culture vaccines are required to stimulate optimal immune responses. Furthermore, current tissue culture vaccines are expensive; thus most people in need of vaccinations (in developing countries) cannot afford them. Hence, there is a need to develop more efficacious and affordable rabies virus vaccines. We propose to develop avirulent live rabies virus vaccines by constructing mutant virus with reduced ability to spread in the nervous system (by mutation of the glycoprotein G) and with reduced rate of viral replication (by mutation and/or rearrangement of genes within the rabies virus genome). This will be accomplished by using the state-of-the-art reverse genetics technology. Our proposal is based on recent findings from us as well as others showing the following. 1) Mutation of the phosphorylated serine at 389 of the N to alanine reduced the rate of viral replication by more than five-fold and virus production by more than 10,000 times. 2) Mutation of the G at residue 333 reduced dramatically the virulence and pathogenicity of rabies virus. 3) Rearrangement of the genes within the vesicular stomatitis virus genome resulted in attenuation and enhancement of immune responses. The rationale for constructing mutated or rearranged rabies viruses is that such altered viruses most likely will be further attenuated than currently available attenuated rabies virus (still induce rabies in neonatal animals). If the further attenuated rabies viruses no longer causes diseases in animals at any age and by any route of inoculation, yet remain immunoqenic, they can be developed as modified live rabies vaccines for humans and animals.