Mucosal Vaccines against Gonorrhea

淋病粘膜疫苗

• 批准号: 7764312
• 负责人: Ann E. Jerse
• 金额: $ 37.85万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-25 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7764312
• 关键词: Active Immunization; Adherence; Adjuvant; Affect; Agglutination; Amino Acid Sequence; Antibiotic Resistance; Antibodies; Antibody-mediated protection; Antigenic Variation; Antigens; Attenuated; Binding; Binding Proteins; Biological Assay; Biological Response Modifiers; Cells; Cervical; Cholera Toxin; Collaborations; Complement; Complement Factor H; Cyclic Peptides; Development; Dose; Effectiveness; Epithelial Cells; Female; Genital system; Gonorrhea; HIV; Human; Human Factor H; Immune; Immune response; Immunization; Immunoglobulin A; Immunoglobulin G; Immunoglobulin Variable Region; Immunology; In Vitro; Infection; Infection prevention; Intestines; Ion Channel; Laboratories; Lead; Length; Lipoproteins; Measures; Mediating; Membrane; Membrane Proteins; Modeling; Multi-Drug Resistance; Mus; Neisseria gonorrhoeae; Outer Limiting Membrane; Passive Immunization; Pelvic Inflammatory Disease; Peptides; Play; PorB porin; Predisposition; Protein Binding; Proteins; Recombinants; Relative (related person); Research; Resistance; Role; Secretory Immunoglobulin A; Serum; Sexually Transmitted Diseases; Surface; System; Testing; Vaccination; Vaccine Antigen; Vaccines; Vagina; antimicrobial peptide; bactericide; base; complement 4b-binding protein; design; efflux pump; gonorrhea vaccine; granulocyte; in vivo; interest; mouse model; mucosal vaccine; mutant; novel; porin; prophylactic; protective efficacy; resistant strain; response; tool; transmission process; vaccine candidate

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.

有效的淋病疫苗的开发受到淋病疫苗抗原变异性的挑战 淋病奈瑟氏菌 (Gc) 表面存在，且缺乏与预防淋病奈瑟菌 (Gc) 相关的信息 感染。 Gc 孔蛋白是一种很好的候选疫苗，因为它在外膜 (OM) 中含量丰富， 有限的变异性及其通过与 C4bp 结合在上皮细胞侵袭和血清抵抗中的作用 蛋白质 (C4BP) 和 H 因子 (fH)。其他有希望的疫苗靶点是 MtrE，即 MtrCMtrD 的 OM 通道 MtrE 主动外排泵和 OmpA（一种保守的 OM 蛋白，赋予粘附和侵袭能力） 人类生殖道细胞。我们的主要目标是针对活性中GC孔蛋白的表面暴露环 以及针对淋病的被动免疫策略。我们还将测试MtrE的保护功效 和OmpA。具体来说，我们将 1) 表征针对选定孔蛋白环的体外和体内抗体 保护作用与阻断 C4BP 和 fH 与血清抗性菌株结合的能力相关。这 将针对环孔蛋白肽来测量抗体的杀菌和调理吞噬活性 不同孔蛋白类型的 Gc 菌株。 Abs抑制孔蛋白介导的人宫颈侵袭的能力 将测试细胞和阻断人 fH 和 C4BP 与血清抗性 Gc 的结合。我们将表演 对正常、补体和粒细胞缺陷小鼠进行被动保护研究，以确定 抗体介导的保护机制。我们还将 ii) 制定积极的免疫策略 基于环状孔蛋白环肽来保护小鼠免受 Gc 感染。肽和的最佳剂量 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 决定。将用最有希望的肽对小鼠进行免疫，并免疫和未免疫 小鼠将接受同源 Gc 菌株的阴道内攻击，以确定免疫是否会影响 对感染或定植负荷的易感性。对两种肽的局部和全身免疫反应 显示出保护作用的肽和不具有保护作用的类似肽将被彻底表征以定义 最后，我们将研究 MtrE 和 OmpA 的保护潜力： 候选疫苗基于其在实验性小鼠感染中表现出的重要性。我们将 利用 MtrE 和 OmpA 特异性抗体研究体外保护的相关性并执行 使用纯化的 MtrE、OmpA 和这些分子的表面暴露区域进行免疫/攻击研究 测量它们作为体内淋病奈瑟菌疫苗抗原的潜力。