CARIES IMMUNITY: REGULATION OF ANTI-S. MUTANS RESPONSES

本次更新补助金申请的主要目的是为了更好地 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，诱导唾液伊加抗体， S.变形细胞或部分纯化的产物诱导有效的 在啮齿类动物的龋齿免疫中，我们几乎不了解细胞免疫， 在诱导这种唾液免疫应答中发生的事件。 在 我们的研究，我们将系统地分离和纯化主要的CW 组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，以及细胞表面相关的 proteins. 在这些实验的第一阶段，我们将研究 小鼠和大鼠T和B淋巴细胞对这些抗体的淋巴增殖反应 纯化的成分。 在这方面，我们过去的研究显示， 血清型CHO是鼠B细胞有丝分裂原和多克隆活化剂， 诱导胸腺非依赖性（TI）免疫应答。 康贝特人将以 由血清型CHO刺激的精确B细胞亚群及其用途 该纯化的物质用于研究伊加应答的诱导。 工作 其他人指出，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 和佐剂活性。并确定PG的哪些成分 表现出免疫增强作用。 在进一步的研究中，S.变形杆菌CW蛋白 将通过ID和2D凝胶电泳分离， 使用单克隆抗体鉴定结构抗原。 这些表面 将用单克隆抗体检测全细胞中的蛋白质， 通过免疫荧光和电子 显微镜 主要表面蛋白将通过使用单克隆抗体纯化。 抗体-免疫吸附柱和纯化的蛋白质， 淋巴增生性和胸腺依赖性（TD）和TI免疫应答， 强调伊加反应。 将用以下物质口服免疫生菌大鼠： 适当的S.变形抗原与纯化CW佐剂的组合 并测定唾液sIgA抗体水平。 纯化 诱导显著保护性sIgA抗体的抗原和佐剂 然后将被确定。 这些拟议的研究将大大推进 我们对S.变形杆菌CW组分和 它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.