Anti-curli immunotherapy for bacterial pneumonia

描述(申请人提供)：我们提出了一种新的抗菌策略 针对关键细菌毒力因子CsgA的中和 卷曲菌毛的单体亚基。除了他们的经典角色 在细菌的黏附和附着中，卷曲发挥了毒力因子的作用 它们在诱导诱导型一氧化氮合酶合成中的作用 (INOS)衍生NO。缺乏CsgA的大肠杆菌毒力较低，不能诱导 感染性休克实验模型中的严重低血压。我们的长期合作 目的是推出一种人源性抗CsgA单抗，该单抗可能是 与标准抗生素治疗相结合，用于治疗 有严重的大肠杆菌感染的迹象。我们打算建立 抗CsgA单抗被动免疫的原理证明 致死性大肠杆菌肺炎啮齿动物模型的炎症和死亡率。目标 #1：制备一系列小鼠抗CsgA单抗。小鼠将被 用重组表达的His标记的CsgA免疫。由此产生的 将对产生单抗的杂交瘤进行筛选，以了解其与重组人 CsgA和活的大肠杆菌。20株Ig G级抗CsgA鼠单抗的研制 被选择和克隆。目标2：对一系列药物的体内效力进行排序 鼠源性抗CsgA单抗的筛选 在多步骤筛选过程中。任务1：我们将利用一只LD100小鼠 气管内直接滴注大肠杆菌所致肺炎模型。 单抗将在暴露于致死的大肠杆菌和 生存已定。将在以下基础上选择5个MAB进行进一步研究 免受死亡的伤害。任务2：我们将获得药效学生存 5个领先的抗CsgA单抗候选人的简介。任务3：我们将建立一个 E.亚致死模型的抗炎药效学研究 结肠肺炎。结果测量将包括体重、中性粒细胞 渗透(髓过氧化物酶)、感染负荷(CFU)和脂质过氧化 (丙二醛)。我们希望鉴定出一种鼠源性抗CsgA单抗，它可以提供 对大肠杆菌肺炎致死性和肺脏的有效保护作用 受伤。 建议的商业应用：不可用

DESCRIPTION (provided by applicant): We propose a novel antibacterial strategy directed at the neutralization of a critical bacterial virulence factor, csgA the monomeric subunit of curliated fimbriae. In addition to their classic role in bacterial adhesion and attachment, curli act as a virulence factor by virtue of their role in inducing the synthesis of inducible nitric oxide (NO) synthase (iNOS) derived NO. csgA deficient E. coli are less virulent and fail to induce severe hypotension in experimental models of septic shock. Our long-term objective is to market a human anti-csgA monoclonal antibody (MAb) that may be administered, in conjunction with standard antibiotic therapy, for the indication of severe E. coli infection. We intend to establish proof-of-principle that passive immunization with an anti-csgA MAb reduces inflammation and mortality in a rodent model of lethal E. coli pneumonia. Aim #1: Generate a series of murine anti-csgA monoclonal IgG Ab's. Mice will be immunized with recombinantly expressed His-tagged csgA. The resultant MAb-producing hybridomas will be screened for their affinity to recombinant csgA and to live E. coli. A total of 20 IgG-class anti-csgA murine MAb's will be selected and cloned. Aim #2: Rank order the in vivo potency of a series of murine anti-csgA MAb's. The selection of a lead anti-csgA MAb will be based upon a multi-step screening process. Task #1: we will utilize an LD100 murine model of pneumonia produced by direct intratracheal instillation of E. coli. MAb's will be administered after exposure to a lethal inoculum of E. coli and survival determined. 5 MAb's will be selected for further study on the basis of protection from mortality. Task #2: we will obtain a pharmacodynamic survival profile of the 5 lead anti-csgA MAb candidates. Task #3: we will establish a pharmacodynamic anti-inflammatory profile in a sublethal model of E. colipneumonia. Outcome measures will include body weight, neutrophil infiltration (myeloperoxidase), infectious burden (cfu), and lipid peroxidation (malondialdehyde). We expect to identify a murine anti-csgA MAb that provides potent protection against E. coli pneumonia induced lethality and pulmonary injury. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE