Development of therapeutic antibodies for vaccinia virus

描述（由申请人提供）：为了应对生物恐怖分子可能释放的天花（天花），对高危人群接种牛痘病毒（VV）疫苗是唯一可用的预防性干预措施。然而，疫苗有已知的并发症，特别是在免疫功能低下的宿主、孕妇和婴儿中。在过去的疫苗接种工作中，这种并发症在美国是用牛痘人免疫球蛋白（VIG）治疗的，这些球蛋白是从接种过牛痘的人身上获得的。虽然这种材料有功效，但很少知道免疫球蛋白的哪些成分是有效的，而且功效可能批次之间存在差异。此外，由于常规疫苗接种已中断多年，免疫球蛋白的供应有限，人们对其安全性感到担忧。我们的长期目标是开发一种针对VV蛋白的明确和高亲和力抗体混合物，在需要大规模VV疫苗接种时取代传统的VV抗体。我们的策略是开发针对各种已知在小鼠模型中引起中和和/或保护反应的VV包膜蛋白的单克隆抗体。这些抗体的混合物，我们称之为VIG-R（替代），将提供一种统一和安全的VV免疫治疗试剂来源。

DESCRIPTION (provided by applicant): In response to the possible release of smallpox (variola) by bioterrorists, vaccinia virus (VV) vaccination of an at-risk population is the only available prophylactic intervention. However, the vaccine has known complications, especially in immunocompromised hosts, pregnant women and infants. In past vaccination efforts, such complications were treated in the U.S. with vaccinia human immune globulin (VIG) obtained from VV immunized people. Although this material had efficacy, little is known about which components of the immune globulin were effective and there could be batch-to-batch variation in efficacy. Moreover, since routine vaccination has been discontinued for many years, there is only a limited supply of VIG available, and there are concerns about its safety. Our long-term goal is to develop a cocktail of defined and high affinity antibodies to VV proteins that will replace the use of traditional VIG in the event that mass VV vaccination is needed. Our strategy is to develop monoclonal antibodies to various VV envelope proteins that are known to provoke a neutralizing and/or protective response in a mouse model. A cocktail of such antibodies, which we will call VIG-R (replacement) will provide a uniform and secure source of a VV immune therapeutic reagent. In this application, we propose experiments in four specific aims. These are: 1) to express and characterize VV envelope proteins LIR, A33R and B5R in a baculovirus expression system and to prepare mouse and rabbit antibodies to those proteins; 2) to develop and characterize human monoclonal antibodies to LIR, A33R and BSR produced using phage display; and 3) to test the ability of immune reagents against VV proteins to protect mice from challenge with VV. Fulfillment of the aims of this grant should provide new information about each of the VV proteins and antibodies, as well as a source of reagents that can comprise a VIG-R. If our approach appears promising, we will expand this study to include other VV proteins. The reagents developed in this grant application will also be valuable for basic studies concerning the role of these proteins in the VV life cycle and could themselves be considered as vaccine candidates in future investigations.