MUCOSAL VACCINES AGAINST GONORRHEA

淋病粘膜疫苗

• 批准号: 6511167
• 负责人: MICHAEL W RUSSELL
• 金额: $ 30.89万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-06-01 至 2004-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6511167
• 关键词: Neisseria gonorrhoeae; bacterial antigens; bacterial proteins; gonorrhea vaccine; laboratory mouse; membrane proteins; mucosal immunity; nonhuman therapy evaluation; vaccine development

DESCRIPTION: (Adapted from Applicant's Abstract) The objective of this proposal is to evaluate the use of a newly discovered, highly conserved outer-membrane protein antigen of Neisseria gonorrhoeae, designated NspA, as a potential candidate vaccine against gonorrhea, when administered by mucosal routes designed to induce high levels of antibodies in the genital tract. This will be accomplished by exploiting a novel technology, developed in this laboratory, for fusing bacterial protein antigens to the A2 subunit of cholera toxin (CT) and co-expressing the fusion protein with the nontoxic binding (B) subunit of CT, to form chimeric immunogens of the form NspA-CTA2/B, in which the toxic A1 subunit of CT has been replaced by the desired antigen. Chemical conjugates of NspA and CTB will also be evaluated. Alternative constructs will utilize type II heat-labile enterotoxins of Escherichia coli, which have different binding properties. Immunogens of this type have previously been shown to induce strong mucosa and circulating antibody responses when administered by mucosal routes. Specific IgA and IgG antibody responses in the genital tract (and other mucosal sites) and in the serum will be determined in mice immunized with these constructs as applied by intranasal or intragastric routes. Specific antibody secreting cells, specific T cells and the cytokines secreted by T cells will also be evaluated to assess the immune response in detail. A newly described mouse model of genital tract colonization by N. gonorrhoeae will be used to determine the ability of NspA-CTA2/B chimeric proteins and other constructs to elicit protective immunity against gonococcal infection. Potential mechanisms by which the expected IgA and IgG antibodies to NspA may be effective in protection against gonococcal infection of the genital tract will be examined by developing monoclonal IgA and IgG antibodies from mice mucosally immunized with NspA-CTA2/B constructs, and testing their ability to inhibit gonococcal adherence to and invasion of epithelial cells in culture, and to suppress genital colonization of mice with N. gonorrhoeae. The successful accomplishment of these objectives should provide a basis for further considering NspA as a component of a vaccine against gonorrhea, and for proposing trials designed to evaluate human genital tract immune responses to NspA-CTA2/B chimeric proteins. The information gained about genital tract immunity and the techniques used may also be applicable to other sexually transmitted diseases.

描述：（改编自申请人的摘要）本提案的目标 是评估新发现的、高度保守的外膜的使用 淋病奈瑟氏球菌蛋白抗原，命名为 NspA，作为潜在的 通过粘膜途径施用时的淋病候选疫苗 旨在在生殖道中诱导高水平的抗体。这将是 通过利用该实验室开发的新技术来实现， 用于将细菌蛋白抗原与霍乱毒素 (CT) 的 A2 亚基融合 并与无毒结合（B）亚基共表达融合蛋白 CT，形成NspA-CTA2/B形式的嵌合免疫原，其中有毒的A1 CT 亚基已被所需抗原取代。的化学共轭物 NspA 和 CTB 也将接受评估。替代结构将利用类型 II 大肠杆菌不耐热肠毒素，具有不同的结合力 特性。此类免疫原先前已被证明可诱导强 通过粘膜途径给药时粘膜和循环抗体反应。 生殖道（和其他粘膜）中的特异性 IgA 和 IgG 抗体反应 位点）和血清中的含量将在用这些免疫的小鼠中测定 通过鼻内或胃内途径施用的构建体。特异性抗体 分泌细胞、特异性T细胞以及T细胞分泌的细胞因子 还可以进行评估以详细评估免疫反应。新描述的 淋病奈瑟菌生殖道定植的小鼠模型将用于 确定 NspA-CTA2/B 嵌合蛋白和其他构建体的能力 引发针对淋球菌感染的保护性免疫力。潜在机制 由此预期的 NspA 的 IgA 和 IgG 抗体可能有效 将检查对生殖道淋球菌感染的保护 通过从粘膜免疫小鼠中开发单克隆 IgA 和 IgG 抗体 使用 NspA-CTA2/B 构建体，并测试其抑制淋球菌的能力 培养物中上皮细胞的粘附和侵袭，并抑制 淋病奈瑟菌在小鼠生殖器定植。成功的成就 这些目标应为进一步考虑 NspA 作为一个 淋病疫苗的组成部分，并提出旨在 评估人类生殖道对 NspA-CTA2/B 嵌合蛋白的免疫反应。 获得的有关生殖道免疫的信息和所使用的技术可能会 也适用于其他性传播疾病。