NEUTRALIZING ANTIBODIES AGAINST ORTHOPOX VIRUSES

人们担心，天花病毒是天花的病原体，二十多年前作为人类病原体被根除，但这次作为生物恐怖主义媒介再次对人类构成威胁。 在生物恐怖袭击中使用天花病毒将通过使用许可的活牛痘病毒疫苗来解决。 这种疫苗可能会引起严重的副作用，可以成功地治疗与牛痘免疫球蛋白（VIG）来自超免疫个体。 然而，VIG供应短缺，未来的可用性是不确定的，此外，还存在使用人血制品用于治疗应用的普遍担忧。该提案旨在制备和表征抗牛痘病毒的人单克隆抗体，其可能配制为抗体混合物，将构成VIG的替代品。 我们将分离针对两种感染形式的牛痘病毒（即细胞内成熟病毒（IMV）和细胞外包膜病毒（EEV））的中和抗体。 我们将特别强调分离抗EEV的抗体，因为EEV介导感染的传播，并且是保护性免疫反应所针对的病毒形式。 将在体外和体内研究牛痘病毒的灭活，并旨在设计在暴露前和暴露后免疫保护中提供针对牛痘病毒感染的保护的抗体混合物。 设计的抗体鸡尾酒可能为天花本身提供治疗。 为了检验被动免疫在非人灵长类动物模型中对天花样疾病的免疫预防和免疫治疗的影响，我们将使用猴痘病毒感染的实验模型

There is concern that variola virus, the causative agent of smallpox which was eradicated as a human pathogen more than two decades ago, forms a threat to humans once again, this time as an agent of bioterrorism. The use of variola virus in a bioterrorist attack would be met by the use of the licensed live vaccinia virus vaccine. This vaccine may cause serious side effects which can be successfully treated with vaccinia immune globulin (VIG) derived from hyperimmune individuals. VIG however is in short supply and future availability is uncertain, and in addition suffers from the general concerns of using human blood products for therapeutic applications. This proposal aims to prepare and characterize human monoclonal antibodies against vaccinia virus which, likely formulated as an antibody cocktail, will constitute a replacement for VIG. We will isolate neutralizing antibodies against both infectious forms of vaccinia virus, i.e. intracellular mature virus (IMV) and extracellular enveloped virus (EEV). We will place particular emphasis on isolating antibodies against EEV, as EEV mediates dissemination of infection and is the viral form against which protective immune responses are directed. Inactivation of vaccinia virus will be studied in vitro and in vivo, and will be aimed at designing an antibody cocktail that provides protection against vaccinia virus infection in pre-exposure and post- exposure immunopropylaxis. The antibody cocktail designed may provide a treatment for smallpox itself. To examine the impact of passive immunization in immunoprophylaxis and immunotherapy of a smallpox-like disease in a non-human primate model, we will use an experimental model of monkeypox virus infection