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：旨在诱导免疫性传染病的研究， 包括龋齿在内，提供了有关微生物的宝贵信息 抗原在诱导保护性反应的重要性，粘膜的作用 防御感染中的免疫系统和IGA抗体涉及 由外部分泌物沐浴的表面和涉及的机制 免疫反应的诱导。该项目的总体目标是定义 由重组，无毒的粘膜疫苗的机制 沙门氏菌菌株表达链球菌的克隆基因，带有和 没有佐剂会诱导对克隆抗原的特定免疫反应，该反应 提供长期保护。具体这些研究将：1）确定 沙门氏菌疫苗的持久性和量的影响 在诱导，性质和 免疫反应的记忆。克隆抗原抗体的水平和同型 在口腔或鼻内免疫的动物的血清和外部分泌物中 （在）使用沙门氏菌疫苗的路线，可在短时间内持续存在 将测量宿主，并产生各种克隆的抗原 ELISA确定这些特征对诱导的影响 粘膜免疫反应。保护将在实验模型中进行评估。 沙门氏菌对克隆蛋白的免疫反应的影响将是 以测量抗原特异性增殖的特征，细胞因子产生 由ELISA和ELISPOT分析以及FACS的共刺激分子表达 在系统性和粘膜组织的细胞制备中。 2）确定效果 粘膜佐剂在调节宿主对重组抗原的反应中 突变链球菌。克隆抗原抗体的水平和同型 在血清中的链球菌和口服免疫的动物的分泌物中的链球菌 与嵌合蛋白的途径，由克隆的抗原遗传连接 到表达霍乱毒素（CTB）或沙门氏菌载体疫苗的B亚基 ELISA将测量各种量的嵌合蛋白+/-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表达 共刺激分子和诱导的细胞因子的谱。 3）确定是否 由链球菌的克隆抗原组成的嵌合蛋白更多 比单独诱导保护性免疫反应的每种抗原有效。 唾液和血清中克隆抗原抗体的水平和同型 将在口腔或途中免疫的动物中测量 突变型链球菌抗原的嵌合蛋白诱导较高的唾液IgA 抗体反应和更大的防御感染的保护 链球菌比单独克隆的蛋白质。结果将与 建立沙门氏疫苗输送系统的实用性和 遗传衍生的嵌合蛋白从A的毒力因子的有用性 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。