SYNTHETIC PEPTIDE VACCINES FOR DENTAL CARIES

治疗龋齿的合成肽疫苗

• 批准号: 2700987
• 负责人: MARTIN A TAUBMAN
• 金额: $ 40.35万
• 依托单位: ADA FORSYTH INSTITUTE, INC.
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-05-01 至 2001-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2700987
• 关键词: B lymphocyte; Salmonella; T lymphocyte; active sites; antibody formation; chemical conjugate; chimeric proteins; clinical research; dental caries vaccine; enzyme linked immunosorbent assay; glucans; hexosyltransferase; human subject; immune tolerance /unresponsiveness; immunization; immunoglobulin A; laboratory rat; mucosal immunity; polysaccharides; protein sequence; saliva; synthetic peptide; tetanus toxoid

The overall aim of this proposal is to develop a subunit dental caries vaccine that elicits enhanced immunity to significant peptide and polysaccharide epitopes of the glucosylfransferase(GTF)-glucan virulence pathway that can be delivered in a manner to stimulate protective mucosal immune responses. We have demonstrated that two synthetic peptides, representing areas of GTF function, when injected in a multiple antigenic peptide (MAP) format, protect rats from experimental caries. The research described in this proposal will, in the first specific aim, evaluate the immunogenicity in rats and protective potential of synthetic peptides associated with a putative second catalytic site of GTF administered in a MAP format. Antibody will be determined by ELISA and T cell responses will be evaluated by proliferation assays. The antigenicity in humans and the presence of B and T cell epitopes will also be investigated using overlapping truncated peptides. Also, we will seek a surrogate universal T cell epitope or GTF T cell epitope in the outbred Sprague-Dawley rat. In the second aim, these peptide sequences, together with those already shown to be effective, will permit the design of multiple sequence, multiple epitopic peptide vaccines in a MAP format. These multiple sequence MAPs may by themselves, or by coimmunization with MAP mixtures, achieve enhanced functional inhibition of GTF as judged by 14C glucose incorporation into glucan and protection in the rodent dental caries model. In the third specific aim the conjugation of protein to polysaccharide which can significantly intensify immune responses to both components will be used to evaluate antibody to glucan and to enhance antibody to multiple epitopic sequences. To explore the potential amplification in immunity achieved by this technique, we will conjugate these multiple epitopic peptide sequences directiy to glucan, or to glucan through tetanus toxoid, and evaluate the enhancement in immunological and dental caries-protective effect of the individual and combined components in animal studies. Since we hypothesize that immunity in saliva is likely to result in the most direct protection, in the fourth specific aim we will explore the effect of mucosal application of the subunit vaccine on salivary antibody formation and protection. Subunit vaccines will be delivered to inductive mucosal sites directly in Al(OH)3 Also, attenuated Salmonella constructs will be developed to express tetanus toxin fragment C-GTF peptide chimeric proteins. These attenuated Salmonella encoding GTF peptides to be expressed as protein fusions will also be administered to inductive mucosal sites and evaluated immunologically and for the ability to elicit protection. Our goal is to design a vaccine that contains an array of functional and immunologically relevant epitopes, in a format that will result in a sustained and high level of protection against dental caries and is acceptable for human use.

该提案的总体目标是开发一个亚单位龋齿 疫苗可增强对重要肽的免疫力， 葡糖基转移酶（GTF）-葡聚糖毒力的多糖表位 可以以刺激保护性粘膜的方式传递的途径 免疫反应。我们已经证明了两种合成肽， 当注射到多抗原中时，代表 GTF 功能的区域 肽（MAP）形式，保护大鼠免受实验性龋齿的侵害。研究 本提案中描述的第一个具体目标将评估 大鼠的免疫原性和合成肽的保护潜力 与 GTF 的假定第二个催化位点相关 地图格式。抗体将通过 ELISA 测定，T 细胞反应将 通过增殖测定进行评估。人类的抗原性和 B 细胞和 T 细胞表位的存在也将使用以下方法进行研究 重叠的截短肽。此外，我们将寻求一个通用的替代品 近交系 Sprague-Dawley 大鼠中的 T 细胞表位或 GTF T 细胞表位。在 第二个目标是这些肽序列以及已经显示的肽序列 为了有效，将允许设计多个序列、多个 MAP 格式的表位肽疫苗。这些多序列MAP 可以单独或通过与 MAP 混合物共免疫来实现 根据 14C 葡萄糖判断，GTF 的功能抑制增强 掺入葡聚糖并保护啮齿动物龋齿 模型。在第三个具体目标中，蛋白质与 多糖，可以显着增强对两者的免疫反应 组件将用于评估葡聚糖抗体并增强 多个表位序列的抗体。探索潜力 通过这种技术实现的免疫放大，我们将缀合 这些多个表位肽序列直接针对葡聚糖，或针对葡聚糖 通过破伤风类毒素，并评估免疫学和免疫学的增强 龋齿-单个和组合成分的保护作用 在动物研究中。因为我们假设唾液中的免疫力可能 为了实现最直接的保护，在第四个具体目标中，我们 将探讨粘膜应用亚单位疫苗对 唾液抗体的形成和保护。亚单位疫苗将 直接递送至 Al(OH)3 中的感应粘膜位点，同时减弱 将开发沙门氏菌构建体来表达破伤风毒素片段 C-GTF 肽嵌合蛋白。这些减毒沙门氏菌编码 GTF 待表达为蛋白质融合物的肽也将被施用 诱导粘膜位点并评估免疫学和能力 以引起保护。我们的目标是设计一种疫苗，其中包含 功能和免疫学相关表位阵列，格式为 这将导致持续和高水平的保护 龋齿并且可供人类使用。