Genetically Engineered Oral Vaccines & Caries Immunity

基因工程口服疫苗

• 批准号: 6845650
• 负责人: Suzanne M. Michalek
• 金额: $ 4.17万
• 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/6845650
• 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：旨在诱导对传染病免疫的研究， 包括龋齿，提供了关于微生物的有价值的信息 在诱导保护性反应中重要的抗原，粘膜的作用 免疫系统和IgA抗体在预防感染中的作用 由外部分泌物沐浴的表面，以及参与 诱导免疫反应。该项目的总体目标是定义 由重组无毒疫苗组成的黏膜疫苗的作用机制 表达变形链球菌克隆基因的沙门氏菌 在没有佐剂的情况下，诱导对克隆抗原的特异性免疫反应，这 提供长期保障。具体而言，这些研究将：1)确定 沙门氏菌疫苗株持续期和接种量的影响 变形链球菌克隆化抗原的诱导表达、性质及 免疫反应的记忆。针对克隆抗原的抗体水平和同种类型 经口腔或鼻腔免疫的动物的血清和外分泌物 使用沙门氏菌疫苗的路线，沙门氏菌疫苗在 宿主，并产生不同数量的克隆抗原将被测量 通过酶联免疫吸附试验确定这些特性对诱导的影响。 粘膜免疫反应。保护措施将在实验模型中进行评估。 沙门氏菌对克隆蛋白免疫反应的影响将是 以测量抗原特异性增殖、细胞因子产生为特征的 用ELISA法和ELISPOT法检测共刺激分子，用流式细胞仪检测共刺激分子的表达 在来自全身和粘膜组织的细胞制剂中。2)确定效果 粘膜佐剂对宿主对重组抗原应答的调节作用 变形链球菌。猪瘟病毒克隆化抗原的抗体水平和亚型 口服或口服免疫的动物血清和分泌物中的变形链球菌 以由基因连锁的克隆抗原组成的嵌合蛋白为路线 霍乱毒素B亚单位或沙门氏菌载体疫苗表达 不同量的嵌合蛋白+/-游离型CTB将被用ELISA法测定。 将评估沙门氏菌对CTB佐剂特性的影响 通过评估系统和粘膜组织中的细胞表达 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一种抗原都有效。 唾液和血清中抗克隆抗原抗体的水平和亚型 将在口服或注射免疫的动物身上进行测量，以确定 变形链球菌抗原嵌合蛋白诱导唾液IgA升高 抗体反应和对变种人感染的更好保护 链球菌比每一种克隆的蛋白质都单独。结果将与以下方面相关 建立沙门氏菌疫苗递送系统的实用性和 沙门氏菌毒力因子基因衍生嵌合蛋白的有用性 诱导保护性免疫反应的病原体和佐剂 粘膜病原体，包括与口腔有关的病原体。