NEUTRALIZING ANTIBODIES AGAINST ORTHOPOX VIRUSES

人们担心，二十多年前作为人类病原体被消灭的天花病毒天花病毒再次对人类构成威胁，这次是作为生物恐怖主义的媒介。 在生物恐怖袭击中使用天花病毒可以通过使用获得许可的活牛痘病毒疫苗来应对。 这种疫苗可能会引起严重的副作用，可以使用来自超免疫个体的痘苗免疫球蛋白 (VIG) 成功治疗。 然而，VIG 供应短缺，未来的可用性不确定，而且还受到使用人类血液制品用于治疗应用的普遍担忧。该提案旨在制备和表征针对痘苗病毒的人单克隆抗体，该抗体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传染性痘苗病毒的中和抗体，即细胞内成熟病毒（IMV）和细胞外包膜病毒（EEV）。 我们将特别强调分离针对 EEV 的抗体，因为 EEV 介导感染传播，并且是针对保护性免疫反应的病毒形式。 将在体外和体内研究痘苗病毒的灭活，旨在设计一种抗体混合物，在暴露前和暴露后免疫丙轴中提供针对痘苗病毒感染的保护。 设计的抗体混合物可以为天花本身提供治疗。 为了在非人灵长类动物模型中研究被动免疫对天花样疾病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