CARIES IMMUNITY: REGULATION OF ANTI-S. MUTANS RESPONSES

这项续期拨款申请的主要目的是更好地 了解主细胞壁(CW)的重要免疫生物学效应 和变形链球菌的细胞表面成分。虽然我们和 其他研究表明，诱导唾液中的IgA抗体 变形链球菌细胞或对部分纯化产物诱导有效 啮齿动物的龋病免疫力，我们对细胞免疫几乎一无所知 在诱导唾液免疫反应的过程中发生的事件。在……里面 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)及其组分，以及与细胞表面相关的 蛋白质。在这些实验的第一阶段，我们将检查 小鼠和大鼠T、B淋巴细胞对它们的淋巴增殖反应 提纯的成分。在这方面，我们过去的研究表明， CHO血清型是小鼠B细胞有丝分裂原和多克隆激活剂 诱导胸腺非依赖性(TI)免疫反应。我们将确定 CHO血清型刺激的精确B细胞亚群及其应用 这种纯化的物质用于研究对IgA反应的诱导。的工作 其他人则表示，来自其他细菌的LTA和PG表现出 淋巴增殖特性，我们将评估这两种免疫反应 和变形链球菌LTA的佐剂活性，并测定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凝胶电泳法和主表面分离 用单抗鉴定的结构抗原。这些表面 蛋白质将用克隆抗体在整个细胞中进行检测 免疫荧光法和电子法同时检测单个决定簇 显微镜。主要的表面蛋白将用单抗纯化。 抗体免疫吸附柱和纯化蛋白的检测 淋巴增殖性和胸腺依赖(TD)和TI免疫反应， 强调对免疫球蛋白A的反应。诺生菌大鼠将口服免疫 适当的变形链球菌抗原(S)与纯化的CW佐剂联合使用 并测定唾液中抗SIgA抗体水平。纯净的 可诱导显著保护性SIgA抗体的抗原和佐剂 然后就会确定了。这些拟议的研究将大大推进 我们对变形链球菌CW组分和免疫学特性的了解 他们对有效的龋齿疫苗的贡献。

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.