Genetically Engineered Oral Vaccines & Caries Immunity

基因工程口服疫苗

• 批准号: 6634618
• 负责人: Suzanne M. Michalek
• 金额: $ 30.67万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-02-01 至 2006-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6634618
• 关键词: Salmonella; Salmonella vaccines; Streptococcus mutans; T lymphocyte; bacterial antigens; bacterial vaccines; biotechnology; chimeric proteins; cholera toxin; dental caries vaccine; drug administration rate /duration; enzyme linked immunosorbent assay; flow cytometry; gene expression; immunogenetics; immunomodulators; laboratory mouse; laboratory rat; mucosal immunity; nonhuman therapy evaluation; oral administration; protein purification; tissue /cell culture; vaccine development; vector vaccine

DESCRIPTION: Studies aimed at inducing immunity against infectious diseases, including Dental caries, have provided valuable information on microbial antigens important in inducing protective responses, the role of the mucosal immune system and IgA antibodies in defense against infections involving surfaces bathed by external secretions, and mechanisms involved in the induction of immune responses. The overall goal of this project is to define mechanisms by which mucosal vaccines consisting of recombinant, avirulent Salmonella strains expressing cloned genes of mutans streptococci, with and without adjuvant induce specific immune responses to the cloned antigen, which provide long-term protection. Specifically these studies will: 1) Determine the effect of persistence of the Salmonella vaccine strain and the amount of the expressed cloned antigens of mutans streptococci on the induction, nature and memory of immune response. Levels and isotype of antibodies to cloned antigens in serum and external secretions of animals immunized by the oral or intranasal (IN) route with Salmonella vaccines which persist for short or long times in the host, and which produce various amounts of cloned antigen will be measured by ELISA to determine the effect of these characteristics on the induction of mucosal immune responses. Protection will be assessed in an experimental model. The effect of Salmonella on the immune response to the cloned proteins will be characterized by measuring antigen-specific proliferation, cytokine production by ELISA and ELISPOT assay, and expression of co-stimulatory molecules by FACS in cell preparations from systemic and mucosal tissues. 2) Determine the effect of mucosal adjuvants on modulating host responses to recombinant antigens of mutans streptococci. Levels and isotype of antibodies to cloned antigens of mutans streptococci in serum and secretions of animals immunized by the oral or IN route with chimeric protein consisting of cloned antigens genetically linked to the B subunit of cholera toxin (CTB) or Salmonella vector vaccine expressing various amounts of chimeric proteins +/- free CTB will be measured by ELISA. The effect of the Salmonella on the adjuvant properties of CTB will be assessed by evaluating cells from systemic and mucosal tissues for the expression of co-stimulatory molecules and the profile of cytokines induced. 3) Determine if chimeric proteins consisting of cloned antigens of mutans streptococci are more effective than each antigen alone in inducing protective immune responses. Levels and isotype of antibodies to the cloned antigens in saliva and serum will be measured in animals immunized by the oral or IN route to determine if chimeric proteins of mutans streptococci antigens induce higher salivary IgA antibody responses and greater protection against infection by mutans streptococci than each cloned protein alone. The results will be relevant to establish the practicability of Salmonella vaccine delivery systems and the usefulness of genetically derived chimeric proteins from virulence factors of a pathogen and adjuvants for the induction of protective immune responses against mucosal pathogens including those associated with the oral cavity.

描述：旨在诱导针对传染病的免疫力的研究， 包括龋齿，提供了有关微生物的宝贵信息 抗原在诱导保护性反应中很重要，粘膜的作用 免疫系统和 IgA 抗体可防御感染 被外部分泌物沐浴的表面，以及涉及的机制 诱导免疫反应。该项目的总体目标是定义 粘膜疫苗由重组、无毒力组成的机制 表达变形链球菌克隆基因的沙门氏菌菌株，具有和 无需佐剂即可诱导针对克隆抗原的特异性免疫反应， 提供长期保护。具体来说，这些研究将： 1) 确定 沙门氏菌疫苗株的持久性和用量的影响 表达变形链球菌克隆抗原的诱导、性质和 免疫反应的记忆。克隆抗原抗体的水平和同种型 通过口服或鼻内免疫的动物的血清和外分泌物中 (IN) 沙门氏菌疫苗途径，可在短期或长期持续存在 宿主，并产生不同量的克隆抗原将被测量 通过 ELISA 确定这些特性对诱导的影响 粘膜免疫反应。将在实验模型中评估保护作用。 沙门氏菌对克隆蛋白免疫反应的影响将是 通过测量抗原特异性增殖、细胞因子产生来表征 通过 ELISA 和 ELISPOT 测定，并通过 FACS 表达共刺激分子 来自全身和粘膜组织的细胞制剂。 2）确定效果 粘膜佐剂调节宿主对重组抗原反应的研究 变形链球菌。克隆抗原的抗体水平和同种型 口服或免疫接种动物血清和分泌物中的变形链球菌 IN 途径使用由基因连接的克隆抗原组成的嵌合蛋白 霍乱毒素 (CTB) 或沙门氏菌载体疫苗的 B 亚基表达 将通过 ELISA 测量不同量的嵌合蛋白+/-游离 CTB。 将评估沙门氏菌对 CTB 佐剂特性的影响 通过评估来自全身和粘膜组织的细胞的表达 共刺激分子和诱导的细胞因子谱。 3）确定是否 由变形链球菌克隆抗原组成的嵌合蛋白更多 在诱导保护性免疫反应方面比单独使用每种抗原更有效。 唾液和血清中克隆抗原抗体的水平和同种型 将在通过口服或注射途径免疫的动物中进行测量，以确定是否 变形链球菌抗原的嵌合蛋白诱导更高的唾液 IgA 抗体反应和更好的保护免受变异病毒感染 链球菌比单独的每个克隆蛋白更有效。结果将与 建立沙门氏菌疫苗输送系统的实用性和 从毒力因子遗传衍生的嵌合蛋白的用途 病原体和佐剂用于诱导保护性免疫反应 粘膜病原体，包括与口腔相关的病原体。