Antibody-based vaginal microbicides against gonorrhea

基于抗体（Abs）预防感染和/或疾病的安全性、有效性和长期临床使用，基于抗体的杀微生物剂是预防女性淋病的有吸引力的策略。我们假设，干扰淋病奈瑟菌（GC）逃避宿主先天防御能力的抗体将比单独通过抗体介导的清除机制预防感染的抗体更有效。为了促进针对GC的预防剂的开发，我们开发了GC下生殖道感染的雌性小鼠模型。在这里，我们建议测试三个GC表面分子（MtrE，唾液酸转移酶 (Lst)和不透明（Opa）蛋白），其对小鼠中的GC存活具有可检测的影响， 基于抗体的杀微生物剂的潜在目标。我们还将测试孔蛋白特异性抗体的基础上，在体外强有力的证据，孔蛋白保护补体介导的防御。具体来说，我们将（i）。评估Ab对MtrE（MtrCDE的外膜通道）和FarAB（MtrE外排系统）的表面暴露区域在体外降低GC对抗微生物剂的抗性和预防小鼠实验感染中的有效性，ii.）定义特异于参与补体活化下调的GC孔蛋白环的Ab在体外降低血清抗性和预防实验性鼠感染的潜力iii.）确定抗α-2，3唾液酸转移酶（LsO）的Ab在保护GC免受 体外CMP-NANA依赖性血清抗性和降低小鼠中GC感染性，以及iv.）评估针对GC Opa蛋白的第一半可变和第四保守表面暴露环的Ab的功能活性以及这些Ab预防小鼠中GC感染的能力。将通过测量凝集、杀菌和调理吞噬活性来评估每个目标中靶点特异性Ab的保护潜力;还将检测抗体介导的靶分子功能抑制。Abs与 在一个或多个这些功能性测定中，将测试活性的被动预防实验鼠GC感染的能力。这些研究将使我们能够确定相关的保护，这可能会导致有效的单克隆抗体为基础的杀微生物剂和粘膜疫苗的发展。

Antibody-based microbicides are an attractive strategy for preventing gonorrhea in women based on the Isafety, efficacy and the long-standing clinical use of antibodies (Abs) to prevent infection and, or disease. We hypothesize Abs that interfere with the ability of Neisseria gonorrhoeae (GC) to evade host innate defenses will be more effective than those that prevent infection through antibody-mediated clearance mechanisms alone. To facilitate the development of prophylactic agents against GC, we developed a female mouse model of GC lower genital tract infection. Here we propose to test three GC surface molecules (MtrE, sialyl-transferase (Lst), and Opacity (Opa) proteins) that have a detectable impact on GC survival in mice as potential targets of antibody-based microbicides. We will also test porin-specific Abs based on strong in vitro evidence that porin protects against complement-mediated defenses. Specifically, we will i.) assess the effectiveness of Abs to surface-exposed regions of MtrE, the outer membrane channel of the MtrCDE and FarAB, MtrE efflux systems, in decreasing GC resistance to antimicrobial agents in vitro and in preventing experimental infection in mice, ii.) Define the potential of Abs specific for GC porin loops involved in down-regulation of complement activation to decrease serum resistance in vitro and to prevent experimental murine infection iii.) determine the effectiveness of Abs against alpha-2,3 sialyltransferase (LsO in protecting GC from CMP-NANA-dependent serum resistance in vitro and in reducing GC infectivity in mice, and iv.) assess the functional activities of Abs against the first semivariable and fourth conserved surface exposed loops of GC Opa proteins and the capacity of these Abs to prevent GC infection in mice. The protective potential of Abs specific for the target in each aim will be assessed by measuring agglutination, bactericidal, and opsono-phagocytic activity; antibody-mediated inhibition of target molecule function will also be tested. Abs with activity in one or more of these functional assays will be tested for the capacity to passively prevent lexperimental murine GC infection. These studies will allow us to define correlates of protection, which may lead to the development of effective monoclonal antibody-based microbicides and mucosal vaccines.