Mucosal Vaccines against Gonorrhea

有效的淋病疫苗的开发受到了抗原变异性的挑战。 淋病奈瑟菌（Gc）表面和缺乏相关信息的保护， 感染Gc孔蛋白是一种很好的疫苗候选物，因为它在外膜（OM）中丰富， 有限的变异性，以及其通过与C4 bp结合在上皮细胞侵袭和血清抵抗中的作用 蛋白（C4 BP）和因子H（fH）。其他有希望的疫苗靶点是MtrE，MtrCMtrD的OM通道， MtrE主动外排泵和OmpA，一种保守的OM蛋白，可使粘附和侵袭 人类生殖道细胞。我们的主要目标是靶向GC孔蛋白的表面暴露环， 以及针对淋病的被动免疫策略。我们亦会测试MtrE的保护效能 和Ompa。具体而言，我们将1）表征体外和体内针对所选孔蛋白环的Ab 保护和阻断C4 BP-和fH-与血清抗性菌株结合的能力的相关性。的 针对环孔蛋白肽的Ab的杀菌和调理吞噬活性将针对 不同孔蛋白类型的Gc菌株。Abs对孔蛋白介导的人宫颈癌细胞侵袭的抑制作用 将测试人fH和C4 BP与血清抗性Gc的结合。我们将执行 对正常小鼠、补体缺陷小鼠和粒细胞缺陷小鼠进行被动保护研究， 抗体介导的保护机制。我们还将ii）制定积极的免疫战略 基于环孔蛋白环肽来保护小鼠免受Gc感染。肽的最佳剂量和 诱导具有抗Gc活性的高滴度血清和粘膜抗体所需的佐剂类型将是 测定小鼠将用最有希望的肽进行免疫，以及免疫和未免疫 将用同源Gc菌株阴道内攻击小鼠，以确定免疫是否影响 感染易感性或定殖负荷。对两种肽的局部和全身免疫应答 显示出保护作用的肽和不显示出保护作用的类似肽将被彻底表征以定义 保护的相关性，生病。最后，我们将研究MtrE和OmpA的保护潜力， 疫苗候选物基于它们在实验鼠感染中所表现出的重要性。我们将 利用MtrE-和OmpA-特异性抗体来研究保护的体外相关性， 用纯化的MtrE、OmpA和这些分子的表面暴露区域进行的免疫/攻击研究 以测量它们作为抗N.体内淋病。

The development of an effective gonorrhea yaccine is challenged by the antigenic variability of the Neisseria gonorrhoeae (Gc) surface and a lack of information on the correlates of protection against infection. Gc porin is a good vaccine candidate based on its abundance in the outer membrane (OM), limited variability, and its roles in epithelial cell invasion and serum resistance via binding to C4bp-binding protein (C4BP) and factor H (fH). Other promising vaccine targets are MtrE, the OM channel of the MtrCMtrD- MtrE active efflux pump, and OmpA, a conserved OM protein that confers adherence to and invasion of human genital tract cells. Our primary objective is to target surface-exposed loops of GC porin in active and passive immunization strategies against gonorrhea. We will also test the protective efficacy of MtrE and OmpA. Specifically, we will 1) characterize Abs against selected porin loops for in vitro and in vivo correlates of protection and the capacity to block C4BP- and fH-binding to serum resistant strains. The bactericidal and opsonophagocytic activity of Abs against cyclic porin peptides will be measured against Gc strains of different porin types. The capacity of Abs to inhibit porin-mediated invasion of human cervical cells and to block the binding of human fH and C4BP to serum resistant Gc will be tested. We will perform passive protection studies with normal, complement- and granulocyte-deficient mice to identify mechanisms of antibody-mediated protection. We will also ii) develop an active immunization strategy based on cyclic porin loop peptides to protect mice from Gc infection. The optimum dose of peptide and type of adjuvant needed to induce high titer serum and mucosal antibodies with anti-Gc activity will be determined. Mice will be immunized with the most promising peptides, and immunized and unimmunized mice will be challenged intravaginally with the homologous Gc strain to determine if immunization affects susceptibility to infection or colonization load. The local and systemic immune response to two peptides that show protection and an analogous peptide that does not will be thoroughly characterized to define correlates of protection, ill.) Finally, we will investigate the protective potential of MtrE and OmpA as vaccine candidates based on their demonstrated importance in experimental murine infection. We will utilize MtrE- and OmpA-specific antibodies to investigate in vitro correlates of protection and perform immunization/challenge studies with purified MtrE, OmpA and surface-exposed regions of these molecules to measure their potential as vaccine antigens against N. gonorrhoeae in vivo.