CARIES IMMUNITY: REGULATION OF ANTI-S. MUTANS RESPONSES

龋齿免疫：抗 S 的调节。

• 批准号: 3482759
• 负责人: Jerry R McGhee
• 金额: $ 11.52万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 1979
• 资助国家: 美国
• 起止时间: 1979-05-01 至 1992-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3482759
• 关键词: B lymphocyte; Streptococcus mutans; T lymphocyte; antigen antibody reaction; bacterial antigens; bactericidal immunity; cellular immunity; dental caries inhibitor; gel filtration chromatography; immunomodulators; immunoregulation; ingested fluoride therapy; laboratory mouse; laboratory rabbit; laboratory rat; lipopolysaccharides; lysozyme; macrophage; monocyte; oral bacteria; radiotracer; saliva; secretory immune system; surface antigens; tissue /cell culture; virulence

The major objective of this renewal grant application is to better understand the important immunobiologic effects of the major cell wall (CW) and cell surface components of Streptococcus mutans. Although we and others have shown that induction of salivary IgA antibodies to either whole S. mutans cells or to partially purified products induces an effective caries immunity in rodents, we have almost no understanding of the cellular events which occur in the induction of this salivary immune response. In our studies, we will systematically isolate and purify the major CW components including serotype carbohydrate (CHO), lipoteichoic acid (LTA), peptidoglycan (PG) and its component parts, and cell surface associated proteins. In the first stage of these experiments, we will examine lymphoproliferative responses of murin and rat T and B lymphocytes to these purified components. In this regard, our past studies have shown that serotype CHO is a murine B cell mitogen and polyclonal activator and induces thymic independent (TI) immune responses. We will determine the precise B cell subpopulation which is stimulated by serotype CHO and use this purified substance to investigate induction of IgA responses. Work of others has indicated that LTA and PG from other bacteria exhibit lymphoproliferative properties and we will evaluate both immune responses and adjuvant activity of S. mutans LTA and determine which components of PG exhibit immunopotentiation. In additional studies, S. mutans CW proteins will be isolated by ID and 2D gel electrophoresis and major surface structural antigens identified using monoclonal antibodies. These surface proteins will be examined in the whole cell with monclonal antibodies to individual determinants by both immunofluorescence and electron microscopy. Major surface proteins will be purified by use of monoclonal antibody-immunoadsorbent columns and purified proteins tested for lymphoproliferative and thymic dependent (TD) and TI immune responses, with emphasis on IgA responses. Gnotobiotic rats will be orally immunized with appropriate S. mutans antigen(s) in combination with purified CW adjuvant and levels of salivary sIgA antibodies to antigen determined. Purified antigen and adjuvant which induce significant protective sIgA antibodies will then be determined. These proposed studies will significantly advance our understanding of immunologic properties of S. mutans CW components and their contribution to an effective caries vaccine.

此次续展拨款申请的主要目标是更好地 了解主要细胞壁 (CW) 的重要免疫生物学作用 和变形链球菌的细胞表面成分。 虽然我们和 其他人已经表明，诱导唾液 IgA 抗体 S. mutans 细胞或部分纯化的产品诱导有效 啮齿类动物的龋齿免疫力，我们对细胞几乎一无所知 在诱导这种唾液免疫反应时发生的事件。 在 我们的研究，我们将系统地分离和纯化主要的CW 成分包括血清型碳水化合物（CHO）、脂磷壁酸（LTA）、 肽聚糖（PG）及其组成部分，与细胞表面相关 蛋白质。 在这些实验的第一阶段，我们将检查 鼠和大鼠 T 和 B 淋巴细胞对这些物质的淋巴增殖反应 纯化的成分。 对此，我们过去的研究表明 CHO 血清型是一种鼠 B 细胞有丝分裂原和多克隆激活剂 诱导胸腺独立（TI）免疫反应。 我们将确定 由血清型 CHO 刺激的精确 B 细胞亚群并使用 这种纯化的物质用于研究 IgA 反应的诱导。 的工作 其他人表明来自其他细菌的 LTA 和 PG 表现出 淋巴细胞增殖特性，我们将评估两种免疫反应 和 S. mutans LTA 的佐剂活性，并确定 PG 的哪些成分 表现出免疫增强作用。 在其他研究中，变形链球菌 CW 蛋白 将通过 ID 和 2D 凝胶电泳和主表面进行分离 使用单克隆抗体鉴定结构抗原。 这些表面 将使用单克隆抗体在全细胞中检查蛋白质 通过免疫荧光和电子的个体决定因素 显微镜。 主要表面蛋白将通过使用单克隆纯化 抗体免疫吸附柱和纯化的蛋白质经过测试 淋巴增殖性和胸腺依赖性 (TD) 和 TI 免疫反应， 强调 IgA 反应。 知生大鼠将被口服免疫 适当的变形链球菌抗原与纯化的 CW 佐剂组合 并测定唾液针对抗原的 sIgA 抗体水平。 纯化的 诱导显着保护性 sIgA 抗体的抗原和佐剂 然后将被确定。 这些拟议的研究将显着推进 我们对变形链球菌 CW 成分的免疫学特性的理解以及 他们对有效的龋齿疫苗的贡献。