MUCOSAL VACCINES AGAINST GONORRHEA

描述：(改编自申请者摘要)本提案的目的 是为了评估一种新发现的高度保守的外膜的使用 淋病奈瑟菌蛋白抗原，命名为NspA，作为潜在的 淋病候选疫苗，当通过粘膜途径接种时 目的是在生殖道中诱导高水平的抗体。这将是 通过利用这个实验室开发的一项新技术完成的， 用于将细菌蛋白抗原与霍乱毒素(CT)A2亚单位融合 和无毒结合的(B)亚基共表达融合蛋白 CT，形成NSPA-CTA2/B形式的嵌合免疫原，其中毒素A1 CT的亚单位已被所需的抗原取代。化合物的化学结合物 NSPA和CTB也将接受评估。替代构造将利用类型 不同结合力的不耐热肠毒素 属性。这种类型的免疫原此前已被证明能诱导强烈的 通过粘膜途径给药时的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 部位)和血清中的含量将在用这些抗体免疫的小鼠中测定 通过鼻腔或胃内途径应用的构造物。特异性抗体 分泌细胞、特异性T细胞和T细胞分泌的细胞因子 也要进行评估，以详细评估免疫应答。一种新描述的 淋病奈瑟菌生殖道定植小鼠模型将用于 确定NSPA-CTA2/B嵌合蛋白和其他构建物的能力 激发对淋球菌感染的保护性免疫。潜在机制 预期的抗NSPA的IgA和Ig G抗体可能在 将检查对生殖道淋球菌感染的保护措施。 通过从粘膜免疫的小鼠身上制备单抗IgA和Ig G 与NSPA-CTA2/B构建，并测试其抑制淋球菌的能力 在培养中对上皮细胞的黏附和侵袭，并抑制 淋病奈瑟菌感染小鼠的生殖器定植。成功的成就 这些目标应为进一步考虑将《国家行动纲领》作为 淋病疫苗的组成部分，并提出旨在 评价人类生殖道对NSPA-CTA2/B嵌合蛋白的免疫应答。 获得的有关生殖道免疫的信息和使用的技术可能 也适用于其他性传播疾病。

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binding (B) subunit of CT, to form chimeric immunogens of the form NspA-CTA2/B, in which the toxic A1 subunit of CT has been replaced by the desired antigen.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.