AVIRULENT SALMONELLA SP-10 VACCINE FOR SECRETORY IMMUNITY

用于分泌免疫的无毒沙门氏菌 SP-10 疫苗

• 批准号: 3779291
• 负责人: JOHN HERR
• 金额: --
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3779291
• 关键词: Macaca fascicularis; Salmonella vaccines; active immunization; affinity chromatography; antibody; antigens; cervix; contraceptives; drug design /synthesis /production; drug screening /evaluation; enzyme linked immunosorbent assay; female; fertility immunology; gut associated lymphoid tissue; immunoglobulins; in vitro fertilization; laboratory mouse; laboratory rabbit; mucus; oral contraceptives; ovary; plasma cells; radioimmunoassay; secretion; secretory immune system; sperm; stoichiometry; uterus; vaccines

The primary goal of this research proposal is to develop an oral contraceptive vaccine and to study the ability of this oral vaccine to induce infertility and secretory immunity within the female reproductive tract. The oral recombinant contraceptive vaccine to be developed will consist of avirulent strains of Salmonella which will be genetically engineered to express the human sperm immunogen SP-10. The avirulent strains of Salmonella will induce immunity by stimulating the gut associated lymphoid tissues [GALT] to dispel populations of specific antibody secreting cells which will populate the female reproductive organs. Experiments will be conducted on the cynomolgus monkey, Macca fascicularis. During the initial phase of the work the SP-10 immunogen will be engineered into avirulent strains of Salmonella and the stability and immunogenicity of the constructs evaluated in mice. In parallel with this goal, methods will be developed to measure specific antibody concentrations in the fluids and mucus of macaque ovarian follicles, oviducts, uterus and cervix and to determine the numbers and location of specific antibody secreting cells arising in these organs and in the circulation following parenteral immunization. Having developed methods of measuring immune responses in the monkey female tract, the effect of the oral contraceptive vaccine on secretory immunity within the ovaries, oviducts, uterus, cervix and circulation will be evaluated in monkeys. Most importantly, the anti-fertility potential of the oral contraceptive vaccine will be evaluated. Infertility will be correlated with levels of serum antibody as well as the degree to which secretory immunity was induced within the female tract. Finally, antibodies to SP-10 from serum and reproductive tract secretions will be studied for their ability to inhibit monkey in vitro fertilization. Using affinity purified antibodies, the precise stoichiometry of immunoglobulin required for inhibitory effects will be determined and these amounts compared to antibody levels induced within follicular, oviductal and cervical secretions by oral immunization. The sperm based recombinant oral contraceptive vaccine to be developed in this program will provide fundamental information on oviductal secretory immunity with wide application as a model for understanding contraceptive vaccine action.

本研究计划的主要目标是开发一种口腔 避孕疫苗并研究这种口服疫苗的能力 诱发不孕和女性生殖系统的分泌免疫 道。 即将研发的口服重组避孕疫苗将 由无毒力的沙门氏菌菌株组成，该菌株将在基因上 设计用于表达人类精子免疫原 SP-10。 无毒者 沙门氏菌菌株会通过刺激肠道来诱导免疫力 相关淋巴组织 [GALT] 驱散特定人群 抗体分泌细胞将填充女性生殖细胞 器官。 实验将在食蟹猴 Macca 身上进行 束状肌。 在工作的初始阶段，将设计 SP-10 免疫原 沙门氏菌无毒株的稳定性和免疫原性 在小鼠中评估的构建体。 与这个目标同时，方法 将开发用于测量液体中的特定抗体浓度 以及猕猴卵泡、输卵管、子宫和子宫颈的粘液 确定特异性抗体分泌细胞的数量和位置 产生于这些器官和胃肠外循环后 免疫接种。 开发了测量免疫反应的方法 猴雌道，口服避孕疫苗对 卵巢、输卵管、子宫、子宫颈内的分泌免疫 将在猴子中评估血液循环。 最重要的是，口服避孕药的抗生育潜力 将评估疫苗。 不孕不育与以下水平相关： 血清抗体以及分泌性免疫的程度 在女性阴道内诱发。 最后，血清中的 SP-10 抗体 将研究生殖道分泌物的能力 抑制猴体外受精。 使用亲和纯化的抗体， 抑制作用所需的免疫球蛋白的精确化学计量 将确定这些量与诱导的抗体水平相比 通过口服免疫接种在卵泡、输卵管和宫颈分泌物中。 即将开发的基于精子的重组口服避孕疫苗 该计划将提供有关输卵管分泌的基本信息 免疫力作为理解避孕的模型得到广泛应用 疫苗行动。