Genetically Engineered Oral Vaccines & Caries Immunity

基因工程口服疫苗

• 批准号: 6516438
• 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/6516438
• 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：旨在诱导对感染性疾病的免疫力的研究， 包括龋齿，提供了关于微生物的有价值的信息， 抗原在诱导保护性反应中的重要性，粘膜免疫的作用， 免疫系统和伊加抗体防御感染， 被外部分泌物浸泡的表面，以及 诱导免疫反应。该项目的总体目标是定义 由重组、无毒的 表达变异链球菌克隆基因的沙门氏菌菌株， 在没有佐剂的情况下诱导针对克隆抗原的特异性免疫应答， 提供长期保护。具体而言，这些研究将：1）确定 沙门氏菌疫苗株的持久性的影响和 表达的变形链球菌克隆抗原的诱导、性质和 免疫反应的记忆。克隆抗原的抗体水平和同种型 经口或鼻内免疫动物的血清和外分泌物中 (IN)沙门氏菌疫苗的途径，持续时间短或长， 将测量宿主以及产生不同量克隆抗原的宿主 通过ELISA测定这些特征对诱导 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在 其特征在于测量抗原特异性增殖、细胞因子产生 通过ELISA和ELISPOT测定，并通过FACS表达共刺激分子 在来自全身和粘膜组织的细胞制剂中。2)确定影响 调节宿主对重组抗原的应答 变形链球菌克隆抗原抗体的水平和同种型 口服或口服免疫动物的血清和分泌物中的变形链球菌 IN途径，嵌合蛋白由基因连接的克隆抗原组成 霍乱毒素B亚单位（CT B）或沙门氏菌载体疫苗表达 通过ELISA测量嵌合蛋白+/-游离CTB的各种量。 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 由变异链球菌的克隆抗原组成的嵌合蛋白质更多 在诱导保护性免疫应答方面比单独的每种抗原有效。 唾液和血清中抗克隆抗原抗体的水平和同种型 将在经口或IN途径免疫的动物中进行测量，以确定 变形链球菌抗原的嵌合蛋白诱导更高的唾液伊加 抗体反应和更好的保护，防止感染变异 比单独的每种克隆的蛋白质更容易感染链球菌。结果将与 建立沙门氏菌疫苗输送系统的实用性， 来自病毒毒力因子的遗传衍生嵌合蛋白的用途 病原体和佐剂，用于诱导针对 粘膜病原体，包括与口腔相关的那些。