Hybridoma (Monoclonal Antibody) Core

被动抗体接种是唯一可用于即时免疫的策略。 接触生物武器的个人。此外，毒素特异性抗体可以中和毒素。 药物是卓越的，目前构成了人体内中和毒素的唯一手段。这个 自19世纪90年代Behring和Kitasato发明以来，抗毒素血清在人类治疗中的有效性就已为人所知。 破伤风和白喉毒素的抗血清，并展示了它们的预防和治疗特性。 然而，尽管进行了一个世纪的免疫学研究，我们对抗体的特性知之甚少。 对于抗毒素的效果很重要，而中和毒素的机制在很大程度上 猜想。这项应用的重点是产生四种毒素的抗体：炭疽杆菌毒素 (保护性抗原、致死因子和水肿因子蛋白亚基)和金黄色葡萄球菌肠毒素 目的是产生治疗性抗体，以防止这些制剂的生物攻击。在……里面 此外，我们还将产生西尼罗河病毒(WNV)的中和抗体。美国目前正处于 在西尼罗河病毒流行期间，有人担心这种病毒是故意 在一次生物恐怖主义行动中被带入这个国家。虽然这不太可能，但发病率和死亡率 与西尼罗河病毒相关的疾病仍然是一个重大的公共卫生问题。提出了四个具体目标：1) 产生中和(鼠和人)抗炭疽毒素蛋白组分的单抗，S. 金黄色葡萄球菌肠毒素和西尼罗河病毒包膜(E)蛋白；2)鉴定所需抗体属性 最佳的毒素和病毒中和活性；3)产生非常高亲和力的中和衍生物 抗毒素和抗病毒单抗通过以下途径提高杂交瘤细胞的体细胞高突变率 活化诱导脱氨酶的表达；4)确定抗体介导的机制(S) 毒素和病毒中和。

Passive antibody administration is the only strategy available for conferring immediate immunity to individuals exposed to biological weapons. Furthermore, toxin-specific antibodies are toxin neutralizing agents par excellence and currently constitute the only means of neutralizing toxins in the human host. The efficacy of antitoxin sera in human therapy is known since the 1890s when Behring and Kitasato developed antisera to tetanus and diphtheria toxins and demonstrated their prophylactic and therapeutic properties. However, despite a century of immunological study we know relatively little about what antibody properties are important for antitoxin efficacy and the mechanisms involved in toxin neutralization are largely conjecture. This application is focused on the generation of antibodies to four toxins: B. anthracis toxins (protective antigen, lethal factor, and edema factor protein subunits) and Staphylococcus aureus enterotoxin with the goal of generating therapeutic antibodies to protect against biological attack with these agents. In addition we will generate neutralizing antibodies to West Nile Virus (WNV). The United States is currently in the midst of an unfolding WNV epidemic and there has been some concern that this agent was deliberately introduced into this country in an act of bioterrorism. While this is unlikely, the morbidity and mortality associated with WNV remain a significant public health concern. Four Specific Aims are proposed: 1) To generate neutralizing (murine and human) mononclonal antibodies to anthrax toxin protein components, S. aureus enterotroxin, and WNV envelope (E) protein; 2) To identify the antibody attributes necessary for optimal toxin and viral neutralizing activity; 3) To generate very high affinity derivatives of neutralizing antitoxin and antiviral mAbs by increasing the rate of somatic hypermutation in selected hybridomas through expression of activation induced deaminase (AID); 4) To identify the mechanism(s) of antibody-mediated toxin and virus neutralization.