Ability of Antibody Against Coxiella burnetii LPS to Confer Protective Immunity

本应用的目的是表征抗体(Ab)介导的免疫(AMI)的作用 抗伯氏柯克斯体感染。这是基于我们的观察：1)福尔马林灭活I期疫苗 (PI-V)在BALB/c小鼠中对伯氏弧菌毒力攻击有完全保护作用，但I期 疫苗(PII-V)没有提供显著的保护作用；2)PI-V和PII-V诱导不同的抗体应答，而 细胞因子反应相似；3)I相脂多糖(PI-LPS)可诱导 保护作用类似于PI-V，而II相脂多糖(PII-LPS)没有明显的保护作用；4 被动转移PI-V免疫小鼠的免疫血清可诱导与PI-V相似的保护水平 在幼鼠身上接种疫苗。第一个具体目标是确定急性心肌梗死对伯氏梭菌感染的作用。 工作假说是，抗体在PL-V诱导的预防伯氏梭菌感染中起关键作用 我们将评估IS诱导的对腹膜和腹膜腔内感染伯氏梭菌的保护作用。 气雾剂攻击BALB/c小鼠，气雾剂攻击豚鼠。我们将确定抗体是否是 IS诱导保护的活性成分和确定被动转移抗体的动力学 保护。第二个特定目标将确定关键的保护性抗体亚型和表位靶标。 目前的工作假设是，急性心肌梗死的靶点是内毒素核心和/或O侧链多糖。我们会的 从血清中提取免疫球蛋白，确定IS的保护成分是否为抗体。我们会的 然后确定保护性急性心肌梗死的亚型，提出急性心肌梗死发病机制的假说。这个 AMI识别的表位将使用高纯度的内毒素和通过以下途径识别的内毒素成分定义 免疫和吸收实验。作为一种免费的方法，一组抗伯氏梭菌内毒素 将产生单抗(MAb)，并测试其被动免疫的能力 调职。这些研究将提供设计亚单位疫苗的可行性、概念研究的证据， 我们计划将其作为产品开发建议提交给竞争对手继续申请 WRCE。

The objective of this application is to characterize the role of antibody (Ab)-mediated immunity (AMI against Coxiella burnetii infection. It is based on our observations: 1) formalin-inactivated phase I vaccine (PI-V) generated complete protection against virulent C. burnetii challenge in BALB/c mice, but phase I vaccine (PII-V) did not confer significant protection; 2) PI-V and PII-V elicited different Ab responses, while cytokine responses were similar; 3) phase I lipopolysaccharide (PI-LPS) was able to induce a level o protection similar to PI-V while phase II lipopolysaccharide (PII-LPS) conferred no significant protection; 4 passive transfer of immune serum (IS) from PI-V vaccinated mice induced a level of protection similar to PI-V vaccination in naive mice. The first specific aim is to characterize the role of AMI against C. burnetii infection The working hypothesis is that Abs play a critical role in Pl-V-induced protection against C. burnetii infection We will evaluate the contribution of IS-induced protection against C. burnetii infection in intraperitoneal and aerosol challenged BALB/c mice and aerosol challenged guinea pigs. We will determine whether Abs are the active components of IS-induced protection and define the kinetics of passively transferred antibody protection. The second specific aim will determine the key protective antibody isotypes and epitope targets The working hypothesis is that the target of AMI is LPS core and/or O-side chain polysaccharide. We wil ourify immunoglobulin from serum and establish whether the protective component of IS is antibody. We wil then determine the isotype of protective AMI to develop a hypothesis on the mechanism of AMI. The epitopes recognized by AMI will be defined using highly purified LPS, and LPS components through mmunizations and absorption experiments. As a complimentary approach, a panel of anti-C.burnetii LPS monoclonal antibodies (Mabs) will be generated and tested for the ability to confer immunity by passive transfer. These studies will provide the feasibility, proof of concept research to design a subunit vaccine, which we plan to submit as a product development proposal for the competing continuation application for the WRCE.